Integrin antagonists

ABSTRACT

This invention relates to certain novel compounds and derivatives thereof, their synthesis, and their use as vitronectin receptor antagonists. The vitronectin receptor antagonist compounds of the present invention are αvβ3 antagonists, αvβ5 antagonists or dual αvβ3/αvβ5 antagonists useful for inhibiting bone resorption, treating and preventing osteoporosis, and inhibiting restenosis, diabetic retinopathy, macular degeneration, angiogenesis, atherosclerosis, inflammation and tumor growth.

CROSS-REFERENCE TO RELATED APPLICATIONS

The present invention is related to U.S. provisional applications SerialNo. 60/027,867, filed Oct. 30, 1996, the contents of which are herebyincorporated by reference.

FIELD OF THE INVENTION

The present invention provides novel compounds and derivatives thereof,their synthesis, and their use as vitronectin receptor ligands. Moreparticularly, the compounds of the present invention are αvβ3antagonists, αvβ5 antagonists or dual αvβ3/αvβ5 antagonists useful forinhibiting bone resorption, treating and preventing osteoporosis, andinhibiting vascular restenosis, diabetic retinopathy, maculardegeneration, angiogenesis, atherosclerosis, inflammation and tumorgrowth.

BACKGROUND OF THE INVENTION

This invention relates to compounds for inhibiting bone resorption thatis mediated by the action of a class of cells known as osteoclasts.

Osteoclasts are multinucleated cells of up to 400 μm in diameter thatresorb mineralized tissue, chiefly calcium carbonate and calciumphosphate, in vertebrates. They are actively motile cells that migratealong the surface of bone. They can bind to bone, secrete necessaryacids and proteases and thereby cause the actual resorption ofmineralized tissue from the bone.

More specifically, osteoclasts are believed to exist in at least twophysiological states. In the secretory state, osteoclasts are flat,attach to the bone matrix via a tight attachment zone (sealing zone),become highly polarized, form a ruffled border, and secrete lysosomalenzymes and protons to resorb bone. The adhesion of osteoclasts to bonesurfaces is an important initial step in bone resorption. In themigratory or motile state, the osteoclasts migrate across bone matrixand do not take part in resorption until they attach again to bone.

Integrins are transmembrane, heterodimeric, glycoproteins which interactwith extracellular matrix and are involved in osteoclast attachment,activation and migration. The most abundant integrin in osteoclasts(rat, chicken, mouse and human) is the vitronectin receptor, or αvβ3,thought to interact in bone with matrix proteins that contain the RGDsequence. Antibodies to αvβ3 block bone resorption in vitro indicatingthat this integrin plays a key role in the resorptive process. There isincreasing evidence to suggest that αvβ3 ligands can be used effectivelyto inhibit osteoclast mediated bone resoption in vivo in mammals.

The current major bone diseases of public concern are osteoporosis,hypercalcemia of malignancy, osteopenia due to bone metastases,periodontal disease, hyperparathyroidism, periarticular erosions inrheumatoid arthritis, Paget's disease, immobilization-inducedosteopenia, and glucocorticoid treatment.

All these conditions are characterized by bone loss, resulting from animbalance between bone resorption (breakdown) and bone formation, whichcontinues throughout life at the rate of about 14% per year on theaverage. However, the rate of bone turnover differs from site to site,for example, it is higher in the trabecular bone of the vertebrae andthe alveolar bone in the jaws than in the cortices of the long bones.The potential for bone loss is directly related to turnover and canamount to over 5% per year in vertebrae immediately following menopause,a condition which leads to increased fracture risk.

There are currently 20 million people with detectable fractures of thevertebrae due to osteoporosis in the United States. In addition, thereare 250,000 hip fractures per year attributed to osteoporosis. Thisclinical situation is associated with a 12% mortality rate within thefirst two years, while 30% of the patients require nursing home careafter the fracture.

Individuals suffering from all the conditions listed above would benefitfrom treatment with agents which inhibit bone resorption.

Additionally, αvβ3 ligands have been found to be useful in treatingand/or inhibiting restenosis (recurrence of stenosis after correctivesurgery on the heart valve), atherosclerosis, diabetic retinopathy,macular degeneration and angiogenesis (formation of new blood vessels).Moreover, it has been postulated that the growth of tumors depends on anadequate blood supply, which in turn is dependent on the growth of newvessels into the tumor; thus, inhibition of angiogenesis can cause tumorregression in animal models. (See, Harrison's Principles of InternalMedicine, 12th ed., 1991). αvβ3 antagonists, which inhibit angiogenesis,are therefore useful in the treatment of cancer for inhibiting tumorgrowth. (See e.g., Brooks et al., Cell, 79:1157-1164 (1994)).

Moreover, compounds of this invention can also inhibitneovascularization by acting as antagonists of the integrin receptorαvβ5. A monoclonal antibody for αvβ5 has been shown to inhibitVEGF-induced angiogenesis in rabbit cornea and the chick chorioallantoicmembrane model; M. C. Friedlander, et.al., Science 270, 1500-1502, 1995.Thus, compounds that antagonize αvβ5 are useful for treating andpreventing macular degeneration, diabetic retinopathy, and tumor growth.

In addition, certain compounds of this invention antagonize both theαvβ3 and αvβ5 receptors. These compounds, referred to as "dual αvβ3/αvβ5antagonists," are useful for inhibiting bone resorption, treating andpreventing osteoporosis, and inhibiting vascular restenosis, diabeticretinopathy, macular degeneration, angiogenesis, atherosclerosis,inflammation and tumor growth.

It is an object of the present invention to identify compounds whichbind to the αvβ3 receptor, αvβ5 receptor or both the αvβ3 and αvβ5receptors.

It is a further object of the invention to identify compounds which actas antagonists of the αvβ3 receptor. It is another object of theinvention to identify αvβ3 antagonist compounds which are useful agentsfor inhibiting: bone resorption mediated by osteoclast cells,restenosis, atherosclerosis, inflammation, diabetic retinopathy, maculardegeneration and angiogenesis in animals, preferably mammals, especiallyhumans. Still another object of the invention is to identify αvβ3antagonists which cause tumor regression and/or inhibit tumor growth inanimals.

A further object of the invention is to identify αvβ3 antagonists usefulfor preventing or treating osteoporosis. An additional object of theinvention is to identify αvβ3 antagonists useful for treating cancer.

It has now been found that the compounds of the present invention, αvβ3ligands, are useful for inhibiting bone resorption in mammals. Thus, thecompounds of the present invention are useful for preventing or reducingthe incidence of osteoporosis. Additionally, the αvβ3 ligands of thepresent invention are also useful for treating and/or inhibitingrestenosis, diabetic retinopathy, macular degeneration, atherosclerosisand/or angiogenesis in mammals.

SUMMARY OF THE INVENTION

The present invention provides a compound of the formula ##STR1##wherein X is selected from ##STR2## a 5- or 6-membered monocyclicaromatic or nonaromatic ring system containing 0, 1, 2, 3 or 4heteroatoms selected from N, O or S wherein the 5- or 6-membered ringsystem is either unsubstituted or substituted with R¹ and R², or

a 9- to 10-membered polycyclic ring system, wherein one or more of therings is aromatic, and wherein the polycyclic ring system contains 0, 1,2, 3 or 4 heteroatoms selected from N, O or S, and wherein thepolycyclic ring system is either unsubstituted or substituted with R¹and R² ;

Y is selected from ##STR3## Z is absent or is a 4-11 membered aromaticor nonaromatic mono- or polycyclic ring system containing 0 to 6 doublebonds, and containing 0 to 6 heteroatoms chosen from N, O and S, andwherein the ring system is either unsubstituted or substituted on acarbon or nitrogen atom with one or more groups independently selectedfrom R¹⁴, R¹⁵, R¹⁶ and R¹⁷ ; preferably, Z is not a 6-memberedmonocyclic aromatic ring system;

R¹, R², R³, R⁴, R⁵, R¹¹, R¹², R¹³, R¹⁶ and R¹⁷ are each independentlyselected from

hydrogen, halogen, C₁₋₁₀ alkyl, C₃₋₈ cycloalkyl, aryl, aryl C₁₋₈ alkyl,amino, amino C₁₋₈ alkyl, C₁₋₃ acylamino, C₁₋₃ acylamino C₁₋₈ alkyl, C₁₋₆alkylamino, C₁₋₆ alkylamino-C₁₋₈ alkyl, C₁₋₆ dialkylamino, C₁₋₆dialkylamino C₁₋₈ alkyl, C₁₋₄ alkoxy, C₁₋₄ alkoxy C₁₋₆ alkyl,hydroxycarbonyl, hydroxycarbonyl C₁₋₆ alkyl, C₁₋₃ alkoxycarbonyl, C₁₋₃alkoxycarbonyl C₁₋₆ alkyl, hydroxycarbonyl-C₁₋₆ alkyloxy, hydroxy orhydroxy C₁₋₆ alkyl;

R⁶, R⁷, R¹⁴ and R¹⁵ are each independently selected from

hydrogen,

aryl,

halogen,

aryl-(CH₂)_(p) --,

hydroxyl,

C₁₋₈ alkylcarbonylamino,

aryl C₁₋₅ alkoxy,

C₁₋₅ alkoxycarbonyl,

aminocarbonyl,

C₁₋₈ alkylaminocarbonyl,

C₁₋₆ alkylcarbonyloxy,

C₃₋₈ cycloalkyl,

amino,

C₁₋₆ alkylamino,

amino C₁₋₆ alkyl,

arylaminocarbonyl,

aryl C₁₋₅ alkylaminocarbonyl,

aminocarbonyl,

aminocarbonyl C₁₋₆ alkyl,

hydroxycarbonyl,

hydroxycarbonyl C₁₋₆ alkyl,

C₁₋₈ alkyl, either unsubstituted or substituted, with one or more groupsselected from: halogen, hydroxyl, C₁₋₅ alkylcarbonylamino, aryl C₁₋₅alkoxy, C₁₋₅ alkoxycarbonyl, aminocarbonyl, C₁₋₅ alkylaminocarbonyl,C₁₋₅ alkylcarbonyloxy, C₃₋₈ cycloalkyl, oxo, amino, C₁₋₃ alkylamino,amino C₁₋₃ alkyl, arylaminocarbonyl, aryl C₁₋₅ alkylaminocarbonyl,aminocarbonyl, aminocarbonyl C₁₋₄ alkyl, hydroxycarbonyl, orhydroxycarbonyl C₁₋₅ alkyl,

HC.tbd.C(CH₂)_(r) --

C₁₋₆ alkyl-C.tbd.C(CH₂)_(r) --,

C₃₋₇ cycloalkyl-C.tbd.C(CH₂)_(r) --,

aryl-C.tbd.C(CH₂)_(r) --,

C₁₋₆ alkylaryl-C.tbd.C(CH₂)_(r) --,

H₂ C═CH(CH₂)_(r) --,

C₁₋₆ alkyl-CH═CH(CH₂)_(r) --,

C₃₋₇ cycloalkyl-CH═CH(CH₂)_(r) --,

aryl-CH═CH(CH₂)_(r) --,

C₁₋₆ alkylaryl-CH═CH(CH₂)_(r) --,

C₁₋₆ alkyl-SO₂ (CH₂)_(r) --,

C₁₋₆ alkylaryl-SO₂ (CH₂)_(r) --,

C₁₋₆ alkoxy,

aryl C₁₋₆ alkoxy,

aryl C₁₋₆ alkyl,

C₁₋₆ alkylamino C₁₋₆ alkyl,

arylamino,

arylamino C₁₋₆ alkyl,

aryl C₁₋₆ alkylamino,

aryl C₁₋₆ alkylamino C₁₋₆ alkyl,

arylcarbonyloxy,

aryl C₁₋₆ alkylcarbonyloxy,

C₁₋₆ dialkylamino,

C₁₋₆ dialkylamino C₁₋₆ alkyl,

C₁₋₆ alkylaminocarbonyloxy,

C₁₋₈ alkylsulfonylamino,

C₁₋₈ alkylsulfonylamino C₁₋₆ alkyl,

arylsulfonylamino C₁₋₆ alkyl,

aryl C₁₋₆ alkylsulfonylamino,

aryl C₁₋₆ alkylsulfonylamino C₁₋₆ alkyl,

C₁₋₈ alkoxycarbonylamino,

C₁₋₈ alkoxycarbonylamino C₁₋₈ alkyl,

aryloxycarbonylamino C₁₋₈ alkyl,

aryl C₁₋₈ alkoxycarbonylamino,

aryl C₁₋₈ alkoxycarbonylamino C₁₋₈ alkyl,

C₁₋₈ alkylcarbonylamino,

C₁₋₈ alkylcarbonylamino C₁₋₆ alkyl,

arylcarbonylamino C₁₋₆ alkyl,

aryl C₁₋₆ alkylcarbonylamino,

aryl C₁₋₆ alkylcarbonylamino C₁₋₆ alkyl,

aminocarbonylamino C₁₋₆ alkyl,

C₁₋₈ alkylaminocarbonylamino,

C₁₋₈ alkylaminocarbonylamino C₁₋₆ alkyl,

arylaminocarbonylamino C₁₋₆ alkyl,

aryl C₁₋₈ alkylaminocarbonylamino,

aryl C₁₋₈ alkylaminocarbonylamino C₁₋₆ alkyl,

aminosulfonylamino C₁₋₆ alkyl,

C₁₋₈ alkylaminosulfonylamino,

C₁₋₈ alkylaminosulfonylamino C₁₋₆ alkyl,

arylaminosulfonylamino C₁₋₆ alkyl,

aryl C₁₋₈ alkylaminosulfonylamino,

aryl C₁₋₈ alkylaminosulfonylamino C₁₋₆ alkyl,

C₁₋₆ alkylsulfonyl,

C₁₋₆ alkylsulfonyl C₁₋₆ alkyl,

arylsulfonyl C₁₋₆ alkyl,

aryl C₁₋₆ alkylsulfonyl,

aryl C₁₋₆ alkylsulfonyl C₁₋₆ alkyl,

C₁₋₆ alkylcarbonyl,

C₁₋₆ alkylcarbonyl C₁₋₆ alkyl,

arylcarbonyl C₁₋₆ alkyl,

aryl C₁₋₆ alkylcarbonyl,

aryl C₁₋₆ alkylcarbonyl C₁₋₆ alkyl,

C₁₋₆ alkylthiocarbonylamino,

C₁₋₆ alkylthiocarbonylamino C₁₋₆ alkyl,

arylthiocarbonylamino C₁₋₆ alkyl,

aryl C₁₋₆ alkylthiocarbonylamino,

aryl C₁₋₆ alkylthiocarbonylamino C₁₋₆ alkyl,

C₁₋₈ alkylaminocarbonyl C₁₋₆ alkyl,

arylaminocarbonyl C₁₋₆ alkyl,

aryl C₁₋₈ alkylaminocarbonyl, or

aryl C₁₋₈ alkylaminocarbonyl C₁₋₆ alkyl,

wherein any of the alkyl groups may be unsubstituted or substituted withR¹¹ and R¹² ; and provided that the carbon atom to which R⁶ and R⁷ areattached is itself attached to no more than one heteroatom; or R⁶ andR⁷, or R¹⁴ and R¹⁵ are combined to form oxo, in which case the carbonatom to which R⁶ and R⁷ are attached can itself be attached to more thanone heteroatom;

R⁸ and R⁹ are each independently selected from

hydrogen,

aryl,

halogen,

aryl-(CH₂)_(p) --,

hydroxyl,

C₁₋₈ alkylcarbonylamino,

aryl C₁₋₅ alkoxy,

C₁₋₅ alkoxycarbonyl,

aminocarbonyl,

C₁₋₈ alkylaminocarbonyl,

C₁₋₆ alkylcarbonyloxy,

C₃₋₈ cycloalkyl,

amino,

C₁₋₆ alkylamino,

amino C₁₋₆ alkyl,

arylaminocarbonyl,

aryl C₁₋₅ alkylaminocarbonyl,

aminocarbonyl,

aminocarbonyl C₁₋₆ alkyl,

hydroxycarbonyl,

hydroxycarbonyl C₁₋₆ alkyl,

C₁₋₈ alkyl, either unsubstituted or substituted, with one or more groupsselected from: halogen, hydroxyl, C₁₋₅ alkylcarbonylamino, aryl C₁₋₅alkoxy, C₁₋₅ alkoxycarbonyl, aminocarbonyl, C₁₋₅ alkylaminocarbonyl,C₁₋₅ alkylcarbonyloxy, C₃₋₈ cycloalkyl, oxo, amino, C₁₋₃ alkylamino,amino C₁₋₃ alkyl, arylaminocarbonyl, aryl C₁₋₅ alkylaminocarbonyl,aminocarbonyl, aminocarbonyl C₁₋₄ alkyl, hydroxycarbonyl, orhydroxycarbonyl C₁₋₅ alkyl,

HC.tbd.C(CH₂)_(r) --

C₁₋₆ alkyl-C.tbd.C(CH₂)_(r) --,

C₃₋₇ cycloalkyl-C.tbd.C(CH₂)_(r) --,

aryl-C.tbd.C(CH₂)_(r) --,

C₁₋₆ alkylaryl-C═C(CH₂)_(r) --,

H₂ C═CH(CH₂)_(r) --,

C₁₋₆ alkyl-CH═CH(CH₂)_(r) --,

C₃₋₇ cycloalkyl-CH═CH(CH₂)_(r) --,

aryl-CH═CH(CH₂)_(r) --,

C₁₋₆ alkylaryl-CH═CH(CH₂)_(r) --,

C₁₋₆ alkyl-SO₂ (CH₂)_(r) --,

C₁₋₆ alkylaryl-SO₂ (CH₂)_(r) --,

C₁₋₆ alkoxy,

aryl C₁₋₆ alkoxy,

aryl C₁₋₆ alkyl,

C₁₋₆ alkylamino C₁₋₆ alkyl,

arylamino,

arylamino C₁₋₆ alkyl,

aryl C₁₋₆ alkylamino,

aryl C₁₋₆ alkylamino C₁₋₆ alkyl,

arylcarbonyloxy,

aryl C₁₋₆ alkylcarbonyloxy,

C₁₋₆ dialkylamino,

C₁₋₆ dialkylamino C₁₋₆ alkyl,

C₁₋₆ alkylaminocarbonyloxy,

C₁₋₈ alkylsulfonylamino,

C₁₋₈ alkylsulfonylamino C₁₋₆ alkyl,

arylsulfonylamino C₁₋₆ alkyl,

aryl C₁₋₆ alkylsulfonylamino,

aryl C₁₋₆ alkylsulfonylamino C₁₋₆ alkyl,

C₁₋₈ alkoxycarbonylamino,

C₁₋₈ alkoxycarbonylamino C₁₋₈ alkyl,

aryloxycarbonylamino C₁₋₈ alkyl,

aryl C₁₋₈ alkoxycarbonylamino,

aryl C₁₋₈ alkoxycarbonylamino C₁₋₈ alkyl,

C₁₋₈ alkylcarbonylamino,

C₁₋₈ alkylcarbonylamino C₁₋₆ alkyl,

arylcarbonylamino C₁₋₆ alkyl,

aryl C₁₋₆ alkylcarbonylamino,

aryl C₁₋₆ alkylcarbonylamino C₁₋₆ alkyl,

aminocarbonylamino C₁₋₆ alkyl,

C₁₋₈ alkylaminocarbonylamino,

C₁₋₈ alkylaminocarbonylamino C₁₋₆ alkyl,

arylaminocarbonylamino C₁₋₆ alkyl,

aryl C₁₋₈ alkylaminocarbonylamino,

aryl C₁₋₈ alkylaminocarbonylamino C₁₋₆ alkyl,

aminosulfonylamino C₁₋₆ alkyl,

C₁₋₈ alkylaminosulfonylamino,

C₁₋₈ alkylaminosulfonylamino C₁₋₆ alkyl,

arylaminosulfonylamino C₁₋₆ alkyl,

aryl C₁₋₈ alkylaminosulfonylamino,

aryl C₁₋₈ alkylaminosulfonylamino C₁₋₆ alkyl,

C₁₋₆ alkylsulfonyl,

C₁₋₆ alkylsulfonyl C₁₋₆ alkyl,

arylsulfonyl C₁₋₆ alkyl,

aryl C₁₋₆ alkylsulfonyl,

aryl C₁₋₆ alkylsulfonyl C₁₋₆ alkyl,

C₁₋₆ alkylcarbonyl,

C₁₋₆ alkylcarbonyl C₁₋₆ alkyl,

arylcarbonyl C₁₋₆ alkyl,

aryl C₁₋₆ alkylcarbonyl,

aryl C₁₋₆ alkylcarbonyl C₁₋₆ alkyl,

C₁₋₆ alkylthiocarbonylamino,

C₁₋₆ alkylthiocarbonylamino C₁₋₆ alkyl,

arylthiocarbonylamino C₁₋₆ alkyl,

aryl C₁₋₆ alkylthiocarbonylamino,

aryl C₁₋₆ alkylthiocarbonylamino C₁₋₆ alkyl,

C₁₋₈ alkylaminocarbonyl C₁₋₆ alkyl,

arylaminocarbonyl C₁₋₆ alkyl,

aryl C₁₋₈ alkylaminocarbonyl,

aryl C₁₋₈ alkylaminocarbonyl C₁₋₆ alkyl,

C₇₋₂₀ polycyclyl C₀₋₈ alkylsulfonylamino C₀₋₆ alkyl,

C₇₋₂₀ polycyclyl C₀₋₈ alkylcarbonylamino C₀₋₆ alkyl,

C₇₋₂₀ polycyclyl C₀₋₈ alkylaminosulfonyolamino C₀₋₆ alkyl,

C₇₋₂₀ polycyclyl C₀₋₈ alkylaminocarbonylamino C₀₋₆ alkyl, or

C₇₋₂₀ polycyclyl C₀₋₈ alkyloxycarbonylamino C₀₋₆ alkyl

wherein any of the alkyl groups may be unsubstituted or substituted withR¹¹ and R¹², wherein any of the polycyclyl may be unsubstituted orsubstituted with R¹⁸, R¹⁹, R²⁰ and R²¹, and provided that the carbonatom to which R⁸ and R⁹ are attached is itself attached to no more thanone heteroatom; or R⁸ and R⁹ are combined to form oxo, in which case thecarbon atom to which R⁸ and R⁹ are attached can itself be attached tomore than one heteroatom;

R¹⁰ is selected from

hydrogen,

C₁₋₈ alkyl,

aryl,

aryl C₁₋₈ alkyl,

aryl C₁₋₆ alkoxy,

C₁₋₈ alkylcarbonyloxy C₁₋₄ alkyl,

aryl C₁₋₈ alkylcarbonyloxy C₁₋₄ alkyl,

C₁₋₈ alkylaminocarbonylmethylene, or

C₁₋₈ dialkylaminocarbonylmethylene;

m, n and r are each independently an integer from 0 to 3;

p is an integer from 1 to 4; and

q is an integer from 0 to 2;

and the pharmaceutically acceptable salts thereof.

In one embodiment of the invention is the compound wherein X is

a 9- to 10-membered polycyclic ring system, wherein one or more of therings is aromatic, and wherein the polycyclic ring system contains 0, 1,2, 3 or 4 heteroatoms selected from N, O or S, and wherein thepolycyclic ring system is either unsubstituted or substituted with R¹and R² ; and

all other variables are as defined above;

and the pharmaceutically acceptable salts thereof.

In a class of the invention is the compound of the formula ##STR4##wherein X is selected from ##STR5## Z is absent or is selected from##STR6## R⁶ is selected from hydrogen,

aryl,

--(CH₂)_(p) -aryl,

C₁₋₈ alkyl, either unsubstituted or substituted, with one or more groupsselected from: halogen, hydroxyl, C₁₋₅ alkylcarbonylamino, aryl C₁₋₅alkoxy, C₁₋₅ alkoxycarbonyl, aminocarbonyl, C₁₋₅ alkylaminocarbonyl,C₁₋₅ alkylcarbonyloxy, C₃₋₈ cycloalkyl, oxo, amino, C₁₋₃ alkylamino,amino C₁₋₃ alkyl, arylaminocarbonyl, aryl C₁₋₅ alkylaminocarbonyl,aminocarbonyl, aminocarbonyl C₁₋₄ alkyl, hydroxycarbonyl, orhydroxycarbonyl C₁₋₅ alkyl,

HC.tbd.C(CH₂)_(r) --

C₁₋₆ alkyl-C.tbd.C(CH₂)_(r) --,

C₃₋₇ cycloalkyl-C.tbd.C(CH₂)_(r) --,

aryl-C.tbd.C(CH₂)_(r) --,

C₁₋₆ alkylaryl-C.tbd.C(CH₂)_(r) --,

H₂ C═CH(CH₂)_(r) --,

C₁₋₆ alkyl-CH═CH(CH₂)_(r) --,

C₃₋₇ cycloalkyl-CH═CH(CH₂)_(r) --,

aryl-CH═CH(CH₂)_(r) --,

C₁₋₆ alkylaryl-CH═CH(CH₂)_(r) --,

C₁₋₆ alkyl-SO₂ (CH₂)_(r) --, or

C₁₋₆ alkylaryl-SO₂ (CH₂)_(r) --;

and all other variables are as defined above;

and the pharmaceutically acceptable salts thereof.

In a subclass of the invention is a compound of the formula ##STR7##wherein Y is selected from ##STR8## Z is absent or is selected from##STR9## R⁶ is selected from hydrogen, ##STR10## --(CH₂)p indolyl,HC.tbd.C(CH₂)_(r) --

C₁₋₆ alkyl-C.tbd.C(CH₂)_(r) --,

C₃₋₇ cycloalkyl-C.tbd.C(CH₂)_(r) --,

aryl-C.tbd.C(CH₂)_(r) --,

C₁₋₆ alkylaryl-C.tbd.C(CH₂)_(r) --,

H₂ C═CH(CH₂)_(r) --,

C₁₋₆ alkyl-CH═CH(CH₂)_(r) --,

C₃₋₇ cycloalkyl-CH═CH(CH₂)_(r) --,

aryl-CH═CH(CH₂)_(r) --,

C₁₋₆ alkylaryl-CH═CH(CH₂)_(r) --,

C₁₋₆ alkyl-SO₂ (CH₂)_(r) --, or

C₁₋₆ alkylaryl-SO₂ (CH₂)_(r) --; and

R¹⁰ is selected from hydrogen or C₁₋₈ alkyl; and all other variables areas defined above; and the pharmaceutically acceptable salts thereof

Illustrative of the invention is the compound selected from

Ethyl 3(S)-pyridin-3-yl-3-{2- 3-(5,6,7,8-tetrahydro-1,8!naphthyridin-2-yl)propylcarbamoyl!acetylamino}propionate;

3(S)-pyridin-3-yl-3-{2- 3-(5,6,7,8-tetrahydro-1,8!naphthyridin-2-yl)propylcarbamoyl!acetylamino}propionic acid;

3-(5,6,7,8-Tetrahydro-1,8!naphthyridin-2-ylmethyl)piperidinyl-malonyl-3(S)-pyridin-3-yl-β-alanineethyl ester;

3-(5,6,7,8-Tetrahydro-1,8!naphthyridin-2-ylmethyl)piperidinyl-malonyl-3(S)-pyridin-3-yl-β-alanine;

4-(5,6,7,8-Tetrahydro-1,8!naphthyridin-2-ylmethyl)piperidinyl-malonyl-3(S)-pyridin-3-yl-β-alanineethyl ester;

4-(5,6,7,8-Tetrahydro-1,8!naphthyridin-2-ylmethyl)piperidinyl-malonyl-3(S)-pyridin-3-yl-β-alanine;

4-(5,6,7,8-Tetrahydro-1,8!naphthyridin-2-yl)piperidinyl-malonyl-3(S)-pyridin-3-yl-β-alanineethyl ester; or

4-(5,6,7,8-Tetrahydro-1,8!naphthyridin-2-yl)piperidinyl-malonyl-3(S)-pyridin-3-yl-β-alanine;

and the pharmaceutically acceptable salts thereof

Preferably, the compound is selected from

3(S)-Pyridin-3-yl-3-{2- 3-(5,6,7,8-tetrahydro-1,8!naphthyridin-2-yl)propylcarbamoyl!acetylamino}propionic acid;

3-(5,6,7,8-Tetrahydro-1,8!naphthyridin-2-ylmethyl)piperidinyl-malonyl-3(S)-pyridin-3-yl-β-alanine;

4-(5,6,7,8-Tetrahydro-1,8!naphthyridin-2-ylmethyl)piperidinyl-malonyl-3(S)-pyridin-3-yl-β-alanine;or

4-(5,6,7,8-Tetrahydro-1,8!naphthyridin-2-yl)piperidinyl-malonyl-3(S)-pyridin-3-yl-β-alanine;

and the pharmaceutically acceptable salts thereof

Exemplifying the invention is a pharmaceutical composition comprisingany of the compounds described above and a pharmaceutically acceptablecarrier. An example of the invention is a pharmaceutical compositionmade by combining any of the compounds described above and apharmaceutically acceptable carrier. An illustration of the invention isa process for making a pharmaceutical composition comprising combiningany of the compounds described above and a pharmaceutically acceptablecarrier.

Further illustrating the invention is a method of treating and/orpreventing a condition mediated by antagonism of a vitronectin receptorin a mammal in need thereof, comprising administering to the mammal atherapeutically effective amount of any of the compounds describedabove. Preferably, the condition is selected from bone resorption,osteoporosis, restenosis, diabetic retinopathy, macular degeneration,angiogenesis, atherosclerosis, inflammation, cancer and tumor growth.More preferably, the condition is selected from osteoporosis and cancer.Most preferably, the condition is osteoporosis.

More specifically exemplifying the invention is a method of eliciting avitronectin antagonizing effect in a mammal in need thereof, comprisingadministering to the mammal a therapeutically effective amount of any ofthe compounds or any of the pharmaceutical compositions described above.Preferably, the vitronectin antagonizing effect is an αvβ3 antagonizingeffect; more specifically the αvβ3 antagonizing effect is selected frominhibition of bone resorption, inhibition of restenosis, inhibition ofatherosclerosis, inhibition of angiogenesis, inhibition of diabeticretinopathy, inhibition of macular degeneration, inhibition ofinflammation or inhibition of tumor growth. Most preferably, the αvβ3antagonizing effect is inhibition of bone resorption. Alternatively, thevitronectin antagonizing effect is an αvβ5 antagonizing effect or a dualαvβ3/αvβ5 antagonizing effect. Examples of αvβ5 antagonizing effects areinhibition of: restenosis, atherosclerosis, angiogenesis, diabeticretinopathy, macular degeneration, inflammation or tumor growth.Examples of dual αvβ3/αvβ5 antagonizing effects are inhibition of: boneresorption, restenosis, atherosclerosis, angiogenesis, diabeticretinopathy, macular degeneration, inflammation or tumor growth.

Additional examples of the invention are methods of inhibiting boneresorption and of treating and/or preventing osteoporosis in a mammal inneed thereof, comprising administering to the mammal a therapeuticallyeffective amount of any of the compounds or any of the pharmaceuticalcompositions decribed above.

Further exemplifying the invention is any of the compounds orcompositions described above, further comprising a therapeuticallyeffective amount of a second bone resorption inhibitor; preferably, thesecond bone resorption inhibitor is alendronate.

More particularly illustrating the invention is any of the methods oftreating and/or preventing osteoporosis and/or of inhibiting boneresoption described above, wherein the compound is administered incombination with a second bone resorption inhibitor; preferably, thesecond bone resorption inhibitor is alendronate.

Additional illustrations of the invention are methods of treatinghypercalcemia of malignancy, osteopenia due to bone metastases,periodontal disease, hyperparathyroidism, periarticular erosions inrheumatoid arthritis, Paget's disease, immobilization-inducedosteopenia, and glucocorticoid treatment in a mammal in need thereof,comprising administering to the mammal a therapeutically effectiveamount of any of the compounds or any of the pharmaceutical compositionsdescribed above.

More particularly exemplifying the invention is the use of any of thecompounds described above in the preparation of a medicament for thetreatment and/or prevention of osteoporosis in a mammal in need thereof.Still further exemplifying the invention is the use of any of thecompounds described above in the preparation of a medicament for thetreatment and/or prevention of: bone resorption, tumor growth, cancer,restenosis, artherosclerosis, diabetic retinopathy and/or angiogenesis.

Another illustration of the invention is a drug which is useful fortreating and/or preventing osteoporosis in a mammal in need thereof, theeffective ingredient of the said drug being any of the compoundsdescibed above. More specifically illustrating the invention is a drugwhich is useful for treating and/or preventing: bone resorption, tumorgrowth, cancer, restenosis, artherosclerosis, diabetic retinopathyand/or angiogenesis in a mammal in need thereof, the effectiveingredient of the said drug being any of the compounds described above.

Additional illustrations of the invention are methods of treating tumorgrowth in a mammal in need thereof, comprising administering to themammal a therapeutically effective amount of a compound described aboveand one or more agents known to be cytotoxic or antiproliferative, e.g.,taxol and doxorubicin.

DETAILED DESCRIPTION OF THE INVENTION

Representative compounds of the present invention are avb3 antagonistswhich display submicromolar affinity for the human avb3 receptor.Compounds of this invention are therefore useful for treating mammalssuffering from a bone condition caused or mediated by increased boneresorption, who are in need of such therapy. Pharmacologically effectiveamounts of the compounds, including pharamaceutically acceptable saltsthereof, are administered to the mammal, to inhibit the activity ofmammalian osteoclasts.

The compounds of the present invention are administered in dosageseffective to antagonize the αvβ3 receptor where such treatment isneeded, as, for example, in the prevention or treatment of osteoporosis.For use in medicine, the salts of the compounds of this invention referto non-toxic "pharmaceutically acceptable salts." Other salts may,however, be useful in the preparation of the compounds according to theinvention or of their pharmaceutically acceptable salts. Saltsencompassed within the term "pharmaceutically acceptable salts" refer tonon-toxic salts of the compounds of this invention which are generallyprepared by reacting the free base with a suitable organic or inorganicacid. Representative salts include the following:

Acetate, Benzenesulfonate, Benzoate, Bicarbonate, Bisulfate, Bitartrate,Borate, Bromide, Calcium, Camsylate, Carbonate, Chloride, Clavulanate,Citrate, Dihydrochloride, Edetate, Edisylate, Estolate, Esylate,Fumarate, Gluceptate, Gluconate, Glutamate, Glycollylarsanilate,Hexylresorcinate, Hydrabamine, Hydrobromide, Hydrochloride,Hydroxynaphthoate, Iodide, Isothionate, Lactate, Lactobionate, Laurate,Malate, Maleate, Mandelate, Mesylate, Methylbromide, Methylnitrate,Methylsulfate, Mucate, Napsylate, Nitrate, N-methylglucamine ammoniumsalt, Oleate, Oxalate, Pamoate (Embonate), Palmitate, Pantothenate,Phosphate/diphosphate, Polygalacturonate, Salicylate, Stearate, Sulfate,Subacetate, Succinate, Tannate, Tartrate, Teoclate, Tosylate,Triethiodide and Valerate. Furthermore, where the compounds of theinvention carry an acidic moiety, suitable pharmaceutically acceptablesalts thereof may include alkali metal salts, e.g., sodium or potassiumsalts; alkaline earth metal salts, e.g., calcium or magnesium salts; andsalts formed with suitable organic ligands, e.g., quaternary ammoniumsalts.

The compounds of the present invention, may have chiral centers andoccur as racemates, racemic mixtures and as individual diastereomers, orenantiomers with all isomeric forms being included in the presentinvention. Therefore, where a compound is chiral, the separateenantiomers, substantially free of the other, are included within thescope of the invention; further included are all mixtures of the twoenantiomers. Also included within the scope of the invention arepolymorphs and hydrates of the compounds of the instant invention.

The present invention includes within its scope prodrugs of thecompounds of this invention. In general, such prodrugs will befunctional derivatives of the compounds of this invention which arereadily convertible in vivo into the required compound. Thus, in themethods of treatment of the present invention, the term "administering"shall encompass the treatment of the various conditions described withthe compound specifically disclosed or with a compound which may not bespecifically disclosed, but which converts to the specified compound invivo after administration to the patient. Conventional procedures forthe selection and preparation of suitable prodrug derivatives aredescribed, for example, in "design of prodrugs," ed. H. Bundgaard,Elsevier, 1985. Metabolites of these compounds include active speciesproduced upon introduction of compounds of this invention into thebiological milieu.

The term "therapeutically effective amount" shall mean that amount of adrug or pharmaceutical agent that will elicit the biological or medicalresponse of a tissue, system, animal or human that is being sought by aresearcher or clinician.

The term "bone resorption," as used herein, refers to the process bywhich osteoclasts degrade bone.

The term "alkyl" shall mean straight or branched chain alkanes of one toten total carbon atoms, or any number within this range (i.e., methyl,ethyl, 1-propyl, 2-propyl, n-butyl, s-butyl, t-butyl, etc.).

The term "alkenyl" shall mean straight or branched chain alkenes of twoto ten total carbon atoms, or any number within this range.

The term "alkynyl" shall mean straight or branched chain alkynes of twoto ten total carbon atoms, or any number within this range.

The term "cycloalkyl" shall mean cyclic rings of alkanes of three toeight total carbon atoms, or any number within this range (i.e.,cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl orcyclooctyl).

The term "alkoxy," as used herein, refers to straight or branched chainalkoxides of the number of carbon atoms specified (e.g., C₁₋₅ alkoxy),or any number within this range (i.e., methoxy, ethoxy, etc.).

The term "aryl," as used herein, refers to a monocyclic or polycyclicsystem composed of 5- and 6-membered fully unsaturated or partiallyunsaturated rings, such that the system comprises at least one fullyunsaturated ring, wherein the rings contain 0, 1, 2, 3 or 4 heteroatomschosen from N, O or S, and either unsubstituted or substituted with oneor more groups independently selected from hydrogen, halogen, C₁₋₁₀alkyl, C₃₋₈ cycloalkyl, aryl, aryl C₁₋₈ alkyl, amino, amino C₁₋₈ alkyl,C₁₋₃ acylamino, C₁₋₃ acylamino C₁₋₈ alkyl, C₁₋₆ alkylamino, C₁₋₆alkylamino C₁₋₈ alkyl, C₁₋₆ dialkylamino, C₁₋₆ dialkylamino-C₁₋₈ alkyl,C₁₋₄ alkoxy, C₁₋₄ alkoxy C₁₋₆ alkyl, hydroxycarbonyl, hydroxycarbonylC₁₋₆ alkyl, C₁₋₅ alkoxycarbonyl, C₁₋₃ alkoxycarbonyl C₁₋₆ alkyl,hydroxycarbonyl C₁₋₆ alkyloxy, hydroxy, hydroxy C₁₋₆ alkyl, cyano,trifluoromethyl, oxo or C₁₋₅ alkylcarbonyloxy. Examples of aryl include,but are not limited to, phenyl, naphthyl, pyridyl, pyrazinyl,pyrimidinyl, imidazolyl, benzimidazolyl, indolyl, thienyl, furyl,dihydrobenzofuryl, benzo(1,3) dioxolane, oxazolyl, isoxazolyl andthiazolyl, which are either unsubstituted or substituted with one ormore groups independently selected from hydrogen, halogen, C₁₋₁₀ alkyl,C₃₋₈ cycloalkyl, aryl, aryl C₁₋₈ alkyl, amino, amino C₁₋₈ alkyl, C₁₋₃acylamino, C₁₋₃ acylamino C₁₋₈ alkyl, C₁₋₆ alkylamino, C₁₋₆alkylamino-C₁₋₈ alkyl, C₁₋₆ dialkylamino, C₁₋₆ dialkylamino C₁₋₈ alkyl,C₁₋₄ alkoxy, C₁₋₄ alkoxy C₁₋₆ alkyl, hydroxycarbonyl, hydroxycarbonylC₁₋₆ alkyl, C₁₋₅ alkoxycarbonyl, C₁₋₃ alkoxycarbonyl C₁₋₆ alkyl,hydroxycarbonyl C₁₋₆ alkyloxy, hydroxy, hydroxy C₁₋₆ alkyl, cyano,trifluoromethyl, oxo or C₁₋₅ alkylcarbonyloxy. Preferably, the arylgroup is unsubstituted, mono-, di-, tri- or tetra-substituted with oneto four of the above-named substituents; more preferably, the aryl groupis unsubstituted, mono-, di- or tri-substituted with one to three of theabove-named substituents; most preferably, the aryl group isunsubstituted, mono- or di-substituted with one to two of theabove-named substituents.

Whenever the term "alkyl" or "aryl" or either of their prefix rootsappear in a name of a substituent (e.g., aryl C₀₋₈ alkyl) it shall beinterpreted as including those limitations given above for "alkyl" and"aryl." Designated numbers of carbon atoms (e.g., C₁₋₁₀) shall referindependently to the number of carbon atoms in an alkyl or cyclic alkylmoiety or to the alkyl portion of a larger substituent in which alkylappears as its prefix root.

The terms "arylalkyl" and "alkylaryl" include an alkyl portion wherealkyl is as defined above and to include an aryl portion where aryl isas defined above. The C_(0-m) or C_(1-m) designation where m may be aninteger from 1-10 or 2-10 respectively refers to the alkyl component ofthe arylalkyl or alkylaryl unit. Examples of arylalkyl include, but arenot limited to, benzyl, fluorobenzyl, chlorobenzyl, phenylethyl,phenylpropyl, fluorophenylethyl, chlorophenylethyl, thienylmethyl,thienylethyl, and thienylpropyl. Examples of alkylaryl include, but arenot limited to, toluene, ethylbenzene, propylbenzene, methylpyridine,ethylpyridine, propylpyridine and butylpyridine.

When any substituent includes the definition C₀ (e.g., aryl C₀₋₈ alkyl),the group modified by C₀ is not present in the substituent. Similarly,when any of the variables m, n, q or r is zero, then the group modifiedby the variable is not present; for example, when r is zero, the group"--(CH₂)_(r) C.tbd.CH" is "--C.tbd.CH".

The term "halogen" shall include iodine, bromine, chlorine and fluorine.

The term "oxy" means an oxygen (O) atom. The term "thio" means a sulfur(S) atom. The term "oxo" shall mean=O.

The term "substituted" shall be deemed to include multiple degrees ofsubstitution by a named substitutent. Where multiple substituentmoieties are disclosed or claimed, the substituted compound can beindependently substituted by one or more of the disclosed or claimedsubstituent moieties, singly or plurally.

Under standard nonmenclature used throughout this disclosure, theterminal portion of the designated side chain is described first,followed by the adjacent functionality toward the point of attachment.For example, a C₁₋₅ alkylcarbonylamino C₁₋₆ alkyl substituent isequivalent to ##STR11##

The present invention is also directed to combinations of the compoundsof the present invention with one or more agents useful in theprevention or treatment of osteoporosis. For example, the compounds ofthe instant invention may be effectively administered in combinationwith effective amounts of other agents used in the treatment ofosteoporosis such as bisphosphonate bone resorption inhibitors;preferably, the bone resorption inhibitor is the bisphosphonatealendronate, now sold as FOSAMAX®. Preferred combinations aresimultaneous or alternating treatments of an αvβ3 receptor antagonist ofthe present invention and FOSAMAX®. In accordance with the method of thepresent invention, the individual components of the combination can beadministered separately at different times during the course of therapyor concurrently in divided or single combination forms. The instantinvention is therefore to be understood as embracing all such regimes ofsimultaneous or alternating treatment and the term "administering" is tobe interpreted accordingly. It will be understood that the scope ofcombinations of the compounds of this invention with other agents usefulfor treating αvβ3 related conditions includes in principle anycombination with any pharmaceutical composition useful for treatingosteoporosis.

As used herein, the term "composition" is intended to encompass aproduct comprising the specified ingredients in the specified amounts,as well as any product which results, directly or indirectly, fromcombination of the specified ingredients in the specified amounts.

The compounds of the present invention can be administered in such oraldosage forms as tablets, capsules (each of which includes sustainedrelease or timed release formulations), pills, powders, granules,elixers, tinctures, suspensions, syrups and emulsions. Likewise, theymay also be administered in intravenous (bolus or infusion),intraperitoneal, topical (e.g., ocular eyedrop), subcutaneous,intramuscular or transdermal (e.g., patch) form, all using forms wellknown to those of ordinary skill in the pharmaceutical arts. Aneffective but non-toxic amount of the compound desired can be employedas an αvβ3 inhibitor.

The dosage regimen utilizing the compounds of the present invention isselected in accordance with a variety of factors including type,species, age, weight, sex and medical condition of the patient; theseverity of the condition to be treated; the route of administration;the renal and hepatic function of the patient; and the particularcompound or salt thereof employed. An ordinarily skilled physician,veterinarian or clinician can readily determine and prescribe theeffective amount of the drug required to prevent, counter or arrest theprogress of the condition.

Oral dosages of the present invention, when used for the indicatedeffects, will range between about 0.01 mg per kg of body weight per day(mg/kg/day) to about 100 mg/kg/day, preferably 0.01 to 10 mg/kg/day, andmost preferably 0.1 to 5.0 mg/kg/day. For oral administration, thecompositions are preferably provided in the form of tablets containing0.01, 0.05, 0.1, 0.5, 1.0, 2.5, 5.0, 10.0, 15.0, 25.0, 50.0, 100 and 500milligrams of the active ingredient for the symptomatic adjustment ofthe dosage to the patient to be treated. A medicament typically containsfrom about 0.01 mg to about 500 mg of the active ingredient, preferably,from about 1 mg to about 100 mg of active ingredient. Intravenously, themost preferred doses will range from about 0.1 to about 10 mg/kg/minuteduring a constant rate infusion. Advantageously, compounds of thepresent invention may be administered in a single daily dose, or thetotal daily dosage may be administered in divided doses of two, three orfour times daily. Furthermore, preferred compounds for the presentinvention can be administered in intranasal form via topical use ofsuitable intranasal vehicles, or via transdermal routes, using thoseforms of transdermal skin patches well known to those of ordinary skillin the art. To be administered in the form of a transdermal deliverysystem, the dosage administration will, of course, be continuous ratherthan intermittant throughout the dosage regimen.

In the methods of the present invention, the compounds herein describedin detail can form the active ingredient, and are typically administeredin admixture with suitable pharmaceutical diluents, excipients orcarriers (collectively referred to herein as `carrier` materials)suitably selected with respect to the intended form of administration,that is, oral tablets, capsules, elixirs, syrups and the like, andconsistent with conventional pharmaceutical practices.

For instance, for oral administration in the form of a tablet orcapsule, the active drug component can be combined with an oral,non-toxic, pharmaceutically acceptable, inert carrier such as lactose,starch, sucrose, glucose, methyl cellulose, magnesium stearate,dicalcium phosphate, calcium sulfate, mannitol, sorbitol and the like;for oral administration in liquid form, the oral drug components can becombined with any oral, non-toxic, pharmaceutically acceptable inertcarrier such as ethanol, glycerol, water and the like. Moreover, whendesired or necessary, suitable binders, lubricants, disintegratingagents and coloring agents can also be incorporated into the mixture.Suitable binders include starch, gelatin, natural sugars such as glucoseor beta-lactose, corn sweeteners, natural and synthetic gums such asacacia, tragacanth or sodium alginate, carboxymethylcellulose,polyethylene glycol, waxes and the like. Lubricants used in these dosageforms include sodium oleate, sodium stearate, magnesium stearate, sodiumbenzoate, sodium acetate, sodium chloride and the like. Disintegratorsinclude, without limitation, starch, methyl cellulose, agar, bentonite,xanthan gum and the like.

The compounds of the present invention can also be administered in theform of liposome delivery systems, such as small unilamellar vesicles,large unilamellar vesicles and multilamellar vesicles. Liposomes can beformed from a variety of phospholipids, such as cholesterol,stearylamine or phosphatidylcholines.

Compounds of the present invention may also be delivered by the use ofmonoclonal antibodies as individual carriers to which the compoundmolecules are coupled. The compounds of the present invention may alsobe coupled with soluble polymers as targetable drug carriers. Suchpolymers can include polyvinylpyrrolidone, pyran copolymer,polyhydroxypropylmethacrylamide-phenol,polyhydroxyethylaspartamide-phenol, or polyethyleneoxide-polylysinesubstituted with palmitoyl residues. Furthermore, the compounds of thepresent invention may be coupled to a class of biodegradable polymersuseful in achieving controlled release of a drug, for example,polylactic acid, polyglycolic acid, copolymers of polyactic andpolyglycolic acid, polyepsilon caprolactone, polyhydroxy butyric acid,polyorthoesters, polyacetals, polydihydropyrans, polycyanoacrylates andcrosslinked or amphipathic block copolymers of hydrogels.

In the schemes and examples below, various reagent symbols andabbreviations have the following meanings:

    ______________________________________    AcOH:      Acetic acid.    BH.sub.3.DMS:               Borane.dimethylsulfide.    BOC(Boc):  t-Butyloxycarbonyl.    BOP:       Benzotriazol-1-yloxytris(dimethylamino)-               phosphonium hexafluorophosphate.    CBZ(Cbz):  Carbobenzyloxy or benzyloxycarbonyl.    CDI:       Carbonyldiimidazole.    CH.sub.2 Cl.sub.2 :               Methylene chloride.    CHCl.sub.3 :               Chloroform.    DEAD:      Diethyl azodicarboxylate.    DIAD:      Diisopropyl azodicarboxylate.    DIBAH or   Diisobutylaluminum hydride.    DIBAL-H:    DIPEA:     Diisopropylethylamine.    DMAP:      4-Dimethylaminopyridine.    DME:       1,2-Dimethoxyethane.    DMF:       Dimethylformamide.    DMSO:      Dimethylsulfoxide.    DPFN:      3,5-Dimethyl-1-pyrazolylformamidine nitrate.    DPPA:      Diphenylphosphoryl azide.    EDC:       1-(3-Dimethylaminopropyl)-3-ethylcarbodiimide.    Et:        Ethyl.    EtOAc:     Ethyl acetate.    EtOH:      Ethanol.    HOAc:      Acetic acid.    HOBT:      1-Hydroxybenzotriazole.    LDA:       Lithium diisopropylamide.    MeOH:      Methanol.    NEt.sub.3 :               Triethylamine.    NMM:       N-methylmorpholine.    PCA.HCl:   Pyrazole carboxamidine hydrochloride.    Pd/C:      Palladium on activated carbon catalyst.    Ph:        Phenyl.    pTSA       p-Toluene sulfonic acid.    TEA:       Triethylamine.    TFA:       Trifluoroacetic acid.    THF:       Tetrahydrofuran.    TLC:       Thin Layer Chromatography.    TMEDA:     N,N,N',N'-Tetramethylethylenediamine.    TMS:       Trimethylsilyl.    ______________________________________

The novel compounds of the present invention were prepared according tothe procedure of the following schemes and examples, using appropriatematerials and are further exemplified by the following specificexamples. The most preferred compounds of the invention are any or allof those specifically set forth in these examples. These compounds arenot, however, to be construed as forming the only genus that isconsidered as the invention, and any combination of the compounds ortheir moieties may itself form a genus. The following examples furtherillustrate details for the preparation of the compounds of the presentinvention. Those skilled in the art will readily understand that knownvariations of the conditions and processes of the following preparativeprocedures can be used to prepare these compounds. All temperatures aredegrees Celsius unless otherwise noted.

The following Schemes and Examples describe procedures for makingrepresentative compounds of the present invention. Moreover, byutilizing the procedures described in detail in PCT InternationalApplication Publication Nos. WO95/32710, published Dec. 7, 1995, andWO95/17397, published Jun. 29, 1995, in conjunction with the disclosurecontained herein, one of ordinary skill in the art can readily prepareadditional compounds of the present invention claimed herein.

More specifically, procedures for preparing the N-terminus of thecompounds of the present invention are described in WO 95/32710.Additionally, for a general review describing the synthesis ofβ-alanines which can be utilized as the C-terminus of the compounds ofthe present invention, see Cole, D. C., Recent Stereoselective SyntheticApproaches to b-Amino Acids, Tetrahedron, 1994, 50, 9517-9582; Juaristi,E, et al., Enantioselective Synthesis of β-Amino Acids, AldrichemicaActa, 1994, 27, 3. In particular, synthesis of the 3-methyl β-alanine istaught in Duggan, M. F. et al., J. Med. Chem., 1995, 38, 3332-3341; the3-ethynyl β-alanine is taught in Zablocki, J. A., et al., J. Med. Chem.,1995, 38, 2378-2394; the 3-pyrid-3-yl β-alanine is taught in Rico, J. G.et al., J. Org. Chem., 1993, 58, 7948-7951; and the 2-amino and2-toslylamino β-alanines are taught in Xue, C-B, et al., Biorg. Med.Chem. Letts., 1996, 6, 339-344. ##STR12##

Ethyl 2-(ethylenedioxy)-pentanoate(1-2)

A solution of 1-1 (40.0 g, 250 mmol; Aldrich), ethylene glycol (17.2 g,275 mmol), PTSA (10 mg), and benzene (400 mL) was refluxed for 20 h in aflask equipped with a Dean-Stark trap. The cooled reaction mixture wasconcentrated to give 1-2 as a yellow oil. TLC Rf=0.32 (EtOAc).

¹ H NMR (300 MHz, CDCl₃) δ4.13 (q, J=7 Hz, 2H), 3.94 (m, 4H), 2.32 (m,2H), 1.70 (m, 4H), 1.32 (s, 3H), 1.24 (t, J=7 Hz, 3H).

2-Ethylenedioxy-pentanoic acid (1-3)

A solution of 1-2 (2.0 g, 9.9 mmol), 1N NaOH (24.7 mL, 24.7 mmol), andethanol (49 mL) was stirred at ambient temperature for 6 h followed byconcentration. The residue was diluted with H₂ O and EtOAc and thenacidified with 10% KHSO₄. The organic phase was then washed with brine,dried (MgSO₄), and concentrated to give 1-3 as a colorless oil.

¹ H NMR (300 MHz, CDCl₃) δ3.94 (m, 4H), 2.40 (m, 2H), 1.72 (m, 4H), 1.33(s, 3H).

2-Ethylenedioxy-5-benzyloxycarbonylaminopentane(1-4)

A solution of 1-3 (1.66 g, 9.5 mmol), triethylamine (2.0 mL, 14.3 mmol),diphenylphosphorylazide (2.1 mL, 9.5 mmol), and benzene (48 mL) washeated at reflux for 2 h followed by addition of benzyl alcohol (1.5 mL,14.3 mL). After 20 h, the cooled reaction mixture was diluted with EtOAcand washed with H₂ O, 10% KHSO₄, Sat. NaHCO₃, and brine, dried (MgSO₄)and concentrated. Flash chromatography (silica, 35% EtOAc/hexanes) gave1-4 as a colorless oil.

TLC Rf=0.41 (35% EtOAc/hexanes). ¹ H NMR (300 MHz, CDCl₃) δ7.36 (m, 5H),5.09 (s, 2H), 4.89 (bs, 1H), 3.92 (m, 4H), 3.20 (m, 2H), 1.62 (m, 4H),1.30 (s, 3H).

2-Oxo-5-Benzyloxycarbonylaminopentane (1-5)

A solution of 14 (1.4 g, 5.0 mmol), pTSA (47 mg, 0.25 mmol), and acetone(25 mL) was refluxed for 5 h. The cooled reaction mixture was treatedwith sat. NaHCO₃ (10 mL) and then concentrated. The residue wasdissolved in EtOAc and washed with sat. NaHCO₃, H₂ O, and brine, dried(MgSO₄), and concentrated to furnish 1-5 as a yellow oil.

TLC Rf=0.31 (silica, 30% EtOAc/hexanes). ¹ H NMR (300 MHz, CDCl₃) δ7.32(m, 5H), 5.10 (s, 2H), 4.90 (bs, 1H), 3.20 (m, 2), 2.49 (m, 2), 2.13 (s,3H), 1.69 (m, 2H).

1-Benzyloxycarbonylamino3-( 1,8!naphthyridin-2-yl)-propane (1-7)

A mixture of 1-5 (0.6 g, 2.51 mmol), 1-6 (0.3 g, 2.5 nmol; forpreparation see Het., 1993, 36, 2513) and L-proline (0.0.87 g, 0.75mmol) in absolute ethanol (13 mL) was heated at reflux for 20 h.Following evaporative removal of the solvent, the residue waschromatographed on silica gel, eluting with 0-5% isopropanol/EtOAc togive 1-7 as a yellow oil.

TLCRf=0.20 (ethyl acetate). ¹ H NMR (300 MHz, CHCl₃) δ9.08 (m, 1H), 8.16(m, 1H), 8.10 (d, J=8 Hz, 1H), 7.50-7.30 (m, 8H), 5.08 (m, 3H), 3.33 (m,2H), 3.11 (m, 2H), 2.15 (m, 2H).

3-(5,6,7,8-Tetrahydro- 1,8!naphthyridin-2-yl)propylamine (1-8)

A mixture of 1-7 (0.40 g, 1.24 mmol) and 10% Pd/carbon (0.08 g) in EtOH(12 mL) was stirred under a balloon of hydrogen for 20 h. Filtrationthrough a celite pad and then evaporative removal of the filtratesolvent gave 1-8 as a colorless oil.

TLC Rf=0.47 (silica, 10:1:1 ethanol/NH₄ OH/H₂ O).

¹ H NMR (300 MHz, CD₃ OD) δ7.12 (d, J=7 Hz, 1H), 6.48 (d, J=7 Hz, 1H),3.37 (m, 2H), 2.75-2.50 (m, 6H), 1.70-1.90 (m, 4H),

Ethyl 2- 3-(5,6,7,8-tetrahydro-1,8!naphthyridin-2-yl)-propylcarbamoyl!-acetate (1-9)

A mixture of 1-8 (0.068 g, 0.36 mmol), ethylmalonic acid (0.052 g, 0.40mmol), BOP (0.24 g, 0.54 mmol), and NMM (0.16 mL, 1.4 mmol) in DMF (1.8mL) was stirred at ambient temperature for 20 h. The mixture was dilutedwith ethyl acetate, washed with water, brine, and dried over MgSO₄.Following evaporative removal of the solvent, the residue waschromatographed on silica gel, eluting with 15% ethanol/EtOAc to give1-9 as a colorless glass.

TLC Rf=0.26 (silica, 15% ethanol/EtOAc), ¹ H NMR (300 MHz, CD₃ OD) δ7.12(d, J=7 Hz, 1H), 6.47 (d, J=7 Hz, 1H), 4.17 (q, J=7 Hz, 2H), 3.37 (m,2H), 3.20 (m, 4H), 2.70 (m, 2H), 2.54 (m, 2H), 1.86 (m, 4H), 1.24 (t,J=7 Hz, 3H).

2- 3-(5,6,7,8-Tetrahydro- 1,8!naphthyridin-2-yl)-propylcarbamoyl!-aceticacid (1-10)

To a solution of 1-9 (0.072 g, 0.24 mmol) in EtOH (1.2 mL) was added 1NNaOH (0.6 ml, 0.6 mmol). After stirring for 2 h, the reaction mixturewas neutralized with 1N HCl (0.6 mL) and then concentrated to give 1-10and NaCl.

TLC Rf=0.25 (silica, 3:1 20:1:1 ethanol:NH₄ OH:H₂ O/EtOAc), ¹ H NMR (300MHz, CD₃ OD) δ7.53 (d, J=7 Hz, 1H), 6.60 (d, J=7 Hz, 1H), 3.48 (m, 2H),3.20 (m, 4H), 2.79 (m, 4H), 1.92 (m, 4H).

Ethyl 3(S)-pyridin-3-yl -3-{2- 3-(5,6,7,8-tetrahydro-1,8!naphthyridin-2-yl)-propylcarbamoyl!-acetylamino}propionate (1-12)

A mixture of 1-10 (0.072 g, 0.24 mmol), 1-11 (0.064 g, 0.24 mmol), BOP(0.16 g, 0.36 mmol), and NMM (0.13 mL, 1.2 mmol) in DMF (2 mL) wasstirred at ambient temperature for 20 h. The mixture was diluted withethyl acetate, washed with water, brine, and dried over MgSO₄. Followingevaporative removal of the solvent, the residue was chromatographed onsilica gel, eluting with 1:3 20:1:1 ethanol:NH₄ OH:H₂ O/EtOAc to give1-12 as a colorless glass.

TLC Rf=0.64 (silica, 1:3 20:1:1 ethanol:NH₄ OH:H₂ O/EtOAc), ¹ H NMR (300MHz, CHCl₃) δ8.61 (m, 1H), 8.51 (m, 1H), 8.29 (d, J=8 Hz, 1H), 7.85 (m,1H),7.65 (m, 1H), 7.23 (m, 1H), 7.05 (d, J=7 Hz, 1H), 6.32 (d, J=7 Hz,1H), 5.47 (m, 1H), 5.23 (bs, 1H), 4.09 (q, J=7 Hz, 2H), 3.5-2.5 (m,12H), 1.88 (m, 4H), 1.18 (t, J=7 Hz, 3H).

3(S)-Pyridin-3-yl -3-{2- 3-(5,6,7,8-tetrahydro-1,8!naphthyridin-2-yl)propylcarbamoyl!-acetylamino}propionic acidbis(trifluoracetate) (1-13)

To a solution of 1-12 (0.050 g, 0.11 mmol) in EtOH (1 mL) was added 1NNaOH (0.27 ml, 0.27 mmol). After stirring for 2 h, the reaction mixturewas neutralized with 1N HCl and the solvents evaporated. Prep HPLC (95:5to 50:50 H₂ O/CH₃ CN(0.1% TFA) over 60 min gave 1-13 as a colorless oil.

¹ H NMR (300 MHz, D₂ O) δ8.85 (m, 1H), 8.72 (d, J=8 Hz, 1H), 8.64 (d,J=8 Hz, 1H), 8.08 (d, J=8 Hz, 1H), 8.05 (d, J=8 Hz, 1H), 7.05 (d, J=7Hz, 1H), 6.55 (d, J=7 Hz, 1H), 5.48 (t, J=7 Hz, 1H), 3.45 (m, 2H), 3.22(m, 2H), 3.08 (d, J=7Hz, 2H), 2.76 (m, 2H), 2.65 (m, 2H), 1.88 (m, 4H).##STR13##

N-Boc-Piperidin-3-ylacetic acid (2-2)

A mixture of 2-1 (15 g, 87 mmol), AcOH (125 mL), H₂ O (25 mL), and PtO₂(1.5 g) was shaken under a hydrogen atmosphere (55 PSI) on the Parrapparatus for 4 hr. The reaction mixture was filtered through a celitepad and the filtrate concentrated. The residue was suspended inacetonitrile (500 mL) and 1N NaOH was added until the solid dissolvedand the mixture was basic. Boc₂ O (20.8 g, 95 mmol) was added and thereaction mixture stirred overnight. The acetonitrile was evaporated andthe aqueous phase acidified with 10% KHSO₄. Extraction with EtOAcfollowed by washing the organic phase with brine, drying (MgSO₄), andconcentration gave 2-2 as a clear oil.

¹ H NMR (300 MHz, CDCl₃) δ3.90 (m, 1H), 3.80 (m, 1H), 2.82 (m, 1H), 2.70(m, 1H), 2.4-1.2 (m, 7H).

N-Boc-3-(propan-2-one)piperidine (2-3)

A solution of 2-2 (13.4 g, 55 mmol) and ether (500 mL) at 0° C. wastreated dropwise with MeLi (87 mL, 121 mmol; 1.4 M/ether). Afteraddition was complete the cooling bath was removed and the reactionmixture stirred overnight. The reaction mixture was then poured into aice/water mixture and then extracted with ether. The ether extracts werewashed with brine, dried(MgSO₄), and concentrated. Flash chromatography(silica, 30% EtOAc/hexanes) gave 2-3 as a colorless oil.

TLC Rf=0.59 (silica, 30% EtOAc, hexanes), ¹ H NMR (300 MHz, CDCl₃) δ3.80(m, 2H), 2.92 (m, 1H), 2.64 (m, 1H), 2.40 (m, 1H), 2.28 (m, 1H), 2.15(s, 3H), 2.03 (m, 1H), 1.80 (m, 1H), 1.63-1.00 (m, 1H), 1.45 (s, 9H).

N-Boc-3-( 1,8!-Naphthyridin-2-ylmethyl)piperidine (2-4)

A mixture of 2-3 (74 mg, o.31 mmol), 2-3a (38 mg, 0.31 mmol), L-proline(18 mg, 0.15 mmol), and ethanol (4 mL) was refluxed for 20 hr. Thecooled reaction mixture was concentrated and then purified by flashchromatography (silica, EtOAc) to give 24 as a yellow solid.

TLC Rf=0.24 (silica, EtOAc), ¹ H NMR (300 MHz, CDCl₃) δ9.10 (m, 1H),8.18 (dd, J=8 and 2 Hz, 1H), 8.11 (d, J=8 Hz, 1H), 7.46 (m, 1H), 7.38(d, J=8 Hz, 1H), 3.90 (m, 2H), 3.05-2.60 (m, 3H), 2.22 (m, 1H), 1.81 (m,1H), 1.68 (m, 2H), 1.40 (s, 9H), 1.30 (m, 2H).

N-Boc-3-(5,6,7,8-Tetrahydro- 1,8!naphthyridin-2-ylmethyl)-piperidine(2-5)

A mixture of 24 (1.1 g, 3.3 mmol), 10% Pd/C (108 mg), and ethanol (20mL) was stirred under a hydrogen atmosphere (1 atm) for 20 hr. Thereaction mixture was then filtered through a celite pad and the filtrateconcentrated. Flash chromatography (silica, 80% hexanes/EtOAc) gave 2-5as a colorless oil.

TLC Rf=0.21 (silica, EtOAc), ¹ H NMR (300 MHz, CDCl₃) δ7.03 (d, J=8 Hz,1H), 6.32 (d, J=8 Hz, 1H), 4.79 (bs, 1H), 3.90 (m, 2H), 3.40 (m, 2H),2.78 (m, 1H), 2.70 (m, 2H), 2.57 (m, 1H), 2.44 (m, 2H), 1.90-1.00 (m,7H), 1.41 (s, 9H).

3-(5,6,7,8-Tetrahydro- 1,8!naphthyridin-2-ylmethyl)piperidine .2 HCl(2-6)

HCl gas was bubbled through a solution of 2-5 (0.8 g, 2.5 mmol) andEtOAc (15 mL) at 0° C. for 15 min. After an additional 15 min argon waspassed through the solution for 15 min followed by concentration. Theresidue was azeotroped twice with ether to give 2-6 as a yellow solid.

¹ H NMR (300 MHz, CD₃ OD) δ7.60 (d, J=8 Hz, 1H), 6.63 (d, J=8 Hz, 1H),3.52 (m, 2H), 3.36 (m, 2H), 3.00-2,60 (m, 6H), 2.24 (m, 1H), 2.00-1.70(m, 4H), 1.35 (m, 2H).

Ethyl 3-(5,6,7,8-tetrahydro-1,8!naphthyridin-2-ylmethyl)piperidinyl-malonate (2-7)

A mixture of 2-6 (200 mg, 0.66 mmol), ethyl hydrogen malonate (80 mg,0.60 mmol), HOBT (97 mg, 0.80 mmol), NMM (0.53 mL, 5.3 mmol), and DMF (4mL) was treated with EDC (138 mg, 0.80 mmol). After 20 hr, the reactionmixture was diluted with EtOAc and then washed with sat. NaHCO₃, H² O,and brine, dried (MgSO₄), and concentrated. Flash chromatography(silica, 20% methanol/EtOAc) gave impure 2-7 as an oil which was useddirectly in the next step.

TLC Rf=0.62 (silica, 20% methanol/EtOAC)

3-(5,6,7,8-Tetrahydro-1,8!naphthyridin-2-ylmethyl)piperidinyl-malonyl-3(S)-pyridin-3-yl-β-alanineethyl ester (2-8)

A solution of 2-7 (200 mg, 0.6 mmol), 1N NaOH (0.6 mL), and ethanol (3mL) was stirred at ambient temperature for 1 hr. The reaction mixturewas then treated with 1N HCl (0.6 mL) and concentrated to give asemi-solid. The residue was dissolved in DMF (2 mL) and treatedsequentially with 1-11 (150 mg, 0.56 mmol), HOBT (65 mg, 0.48 mmol), NMM(265 uL, 2.4 mmol), and EDC (92 mg, 0.48 mmol). After 72 hr, thereaction mixture was diluted with EtOAc and then washed with sat,NaHCO₃, H₂ O, and brine, dried (MgSO₄), and concentrated. Flashchromatography (silica, 20% methanol/EtOAc) gave 2-8 as a yellowish oil.

TLC Rf=0.13 (silica, 20% methanol/EtOAc), ¹ H NMR (300 MHz, CDCl₃) δ9.29(bt, 1H), 8.60 (m, 1H), 8.50 (m, 1H), 7.70 (m, 1H), 7.23 (m, 1H), 7.03(m, 1H), 6.38 (m, 1H), 5.49 (m, 1H), 4.10 (q, J=7 Hz, 2H), 3.73 (2H),3.40 (m, 4H), 3.20-2.40 (m, 8H), 2.00-1.20 (m, 7H), 1.17 (t, J=7 Hz, 3H)

3-(5,6,7,8-Tetrahydro-1,8!naphthyridin-2-ylmethyl)piperidinyl-malonyl-3(S)-pyridin-3-yl-β-alanine(2-9)

A mixture of 2-8 (20 mg, 40 umol), 1N NaOH (60 uL), and ethanol (2 mL)was stirred at ambient temperature for 3 hr. The reaction mixture wastreated with 1N HCl (60 uL) and then concentrated. Flash chromatography(silica, 25:10:1:1 to 12:10:1:1 EtOAc/ethanol/H₂ O/NH₄ OH) gave 2-9 as awhite solid.

TLC Rf=0.31 (silica, 50% EtOAc/ 10:1:1 ethano/H₂ O/NH₄ OH!), ¹ H NMR(300 MHz, D₂ O) δ8.55 (m, 1H), 8.44 (m, 1H), 7.93 (m, 1H), 7.52 (m, 2H),6.50 (m, 1H), 5.25 (m, 1H), 3.80-3.40 (m, 8H), 3.00-2.40 (m, 8 H),2.00-1.20 (m, 6H). ##STR14##

N-Boc-4-(propan-2-one)piperidine (3-2)

A solution of 3-1 (10 g, 41 mmol) and ether (400 mL) at 0° C. wastreated dropwise with MeLi (66 mL, 92 mmol; 1.4 M/ether). After additionwas complete the cooling bath was removed and the reaction mixturestirred overnight. The reaction mixture was then poured into a ice/watermixture and then extracted with ether. The ether extracts were washedwith brine, dried(MgSO₄), and concentrated to give 3-2 as an oil.

1H NMR (300 MHz, CDCl₃) δ4.08 (m, 2H), 2.72 (bt, J=12 Hz, 2H), 2.36 (d,J=7 Hz, 2H), 2.14 (s, 3H), 1.97 (m, 1H), 1.63 (m, 2H), 1.45 (s, 9H),1.10 (m, 2H).

N-Boc-4-( 1,8!-Naphthyridin-2-ylmethyl)piperidine (3-3)

A mixture of 3-2 (2.5 g, 10.4 mmol), 1-6 (1.3 g, 10.4 mmol), L-proline(0.6 g, 5.2 mmol), and ethanol (70 mL) was refluxed for 20 hr. Thecooled reaction mixture was concentrated and then purified by flashchromatography (silica, EtOAc) to give 3-3 as a yellow solid.

TLC Rf=0.28 (silica, EtOAc), ¹ H NMR (300 MHz, CDCl₃) δ9.10 (m, 1H),8.18 (dd, J=8 and 2 Hz, 1H), 8.11 (d, J=8 Hz, 1H), 7.46 (m, 1H), 7.35(d, J=8 Hz, 1H), 4.08 (m, 2H), 2.97 (d, J=8 Hz, 2H), 2.70 (m, 2H), 2.24(m, 1H), 1.68 (m, 2H), 1.45 (s, 9H), 1.27 (m, 2H).

N-Boc-4-(5,6,7,8-Tetrahydro- 1,8!naphthyridin-2-ylmethyl)-piperidine(3-4)

A mixture of 3-3 (2.1 g, 6.4 mmol), 10% Pd/C (210 mg), and ethanol (32mL) was stirred under a hydrogen atmosphere (1 atm) for 20 hr. Thereaction mixture was then filtered through a celite pad and the filtrateconcentrated. Flash chromatography (silica, 70% hexanes/EtOAc to EtOAc)gave 34 as a colorless oil.

TLC Rf=0.15 (silica, EtOAc), ¹ H NMR (300 MHz, CDCl₃) δ7.03 (d, J=8 Hz,1H), 6.30 (d, J=8 Hz, 1H), 4.75 (bs, 1H), 4.08 (m, 2H), 3.40 (m, 2H),2.69 (m, 3H), 2.44 (d, J=8 Hz, 2H), 1.90-1.00 (m, 8H), 1.43 (s, 9H).

4-(5,6,7,8-Tetrahydro- 1,8!naphthyridin-2-ylmethyl)piperidine .2 HCl(3-5)

A solution of 3-4 (1.6 g, 4.8 mmol) and dioxane (24 mL) at 0oC wastreated with 4M HCl/dioxane (6.0 mL) followed by removal of the coolingbath. After 5 hr the reaction mixture was concentrated to give 3-5 as ayellow solid.

¹ H NMR (300 MHz, CD₃ OD) δ7.60 (d, J=8 Hz, 1H), 6.63 (d, J=8 Hz, 1H),3.52 (m, 2H), 3.40 (m, 2H), 3.0 (bt, 2H), 2.83 (m, 2H), 2.71 (d, J=8 Hz,2), 2.08 (m, 1H), 2.00-1.85 (m, 4H), 1.50 (m, 2H).

Ethyl 4-(5,6,7,8-tetrahydro-1,8!naphthyridin-2-ylmethyl)piperidinyl-malonate (3-6)

A mixture of 3-5 (150 mg, 0.52 mmol), ethyl hydrogen malonate (76 mg,0.57 mmol), HOBT (70 mg, 0.52 mmol), NMM (0.29 mL, 2.1 mmol), and DMF (4mL) was treated with EDC (97 mg, 0.52 mmol). After 20 hr, the reactionmixture was diluted with EtOAc and then washed with sat. NaHCO₃, H₂ O,and brine, dried (MgSO₄), and concentrated. Flash chromatography(silica, 15% methanol/EtOAc) gave impure 3-6 as an oil.

¹ H NMR (300 MHz, CDCl₃) δ7.05 (d, J=8 Hz, 1H), 6.28 (d, J=8 Hz, 1H),4.80 (bs, 1H), 4.58 (m, 1H), 4.20 (q, J=7 Hz, 2H), 3.70 m, 1H), 3.42 (m,4H), 3.03 (m, 1H), 2.70 (m, 2H), 2.57 (m, 1H), 2.45 (d, J=7 Hz, 2H),2.00-1.00 (m, 7H), 1.28 (t, J=7 Hz, 3H).

4-(5,6,7,8-Tetrahydro-1,8!naphthyridin-2-ylmethyl)piperidinyl-malonyl-3(S)-pyridin-3-yl-β-alanineethyl ester (3-7)

A solution of 3-6 (28 mg, 0.08 mmol), 1N NaOH (0.2 mL), THF (1 mL), andH₂ O (1 mL) was stirred at ambient temperature for 1 hr. The reactionmixture was then treated with 1N HCl (0.2 mL) and concentrated to give asemi-solid. The residue was dissolved in DMF (3 mL) and treatedsequentially with 1-11 (24 mg, 0.088 mmol), HOBT (12 mg, 0.088 mmol),NMM (87 uL, 0.63 mmol), and EDC (17 mg, 0.088 mmol). After 20 hr, thereaction mixture was diluted with EtOAc and then washed with sat,NaHCO₃, H₂ O, and brine, dried (MgSO₄), and concentrated. Flashchromatography (silica, 70:25:5 CHCl₃ /methanol/EtOAc) gave 3-7 as ayellowish oil.

¹ H NMR (300 MHz, CDCl3) δ8.97 (d, J=8 Hz, 0.5H), 8.85 (d, J=8 Hz,0.5H), 8.60 (m, 1H), 8.50 (m, 1H), 7.68 (m, 1H), 7.23 (m, 1H), 7.07 (d,J=8 Hz, 1H), 6.28 (m, 1H), 5.49 (m, 1H), 5.18 (m, 1H), 4.60 (d, J=13 Hz,1H), 3.84 (d, J=13 Hz, 1H), 4.10 (q, J=7 Hz, 2H), 3.73 (2H), 3.40 (m,2H), 3.10-2.80 (m, 4H), 2.70 (m, 1H), 2.67 (m, 1H), 2.45 (m, 1H), 1.95(m, 4H), 1.70 (m, 3H), 1.17 (m, 2H), 1.17 (t, J=7 Hz, 3H)

4-(5,6,7,8-Tetrahydro-1,8!naphthyridin-2-ylmethyl)piperidinyl-malonyl-3(S)-pyridin-3-yl-β-alanine(3-8)

A mixture of 3-7 (8.8 mg, 18 umol), 1N NaOH (44 uL), THF (0.2 mL), andH₂ O (0.2 mL) was stirred at ambient temperature for 3 hr. The reactionmixture was treated with 1N HCl (66 uL) and then concentrated. FlashChromatography (silica, 15:10:1:1 to 12:10:1:1 EtOAc/ethanol/H₂ O/NH₄OH) gave 3-8 as a white solid.

TLC Rf=0.15 (silica, 15:10:1:1 EtOAc/ethanol/H₂ O/NH₄ OH), ¹ H NMR (300MHz, CD₃ OD) δ8.58 (m, 1H), 8.42 (m, 1H), 7.86 (m, 1H), 7.42 (m, 2H),6.50 (m, 1H), 5.40 (m, 1H), 4.48 (m, 1H), 3.88 (m, 1H), 3.70-3.00 (m,5H), 3.00-2.40 (m, 7 H), 2.00-1.00 (m, 7H). ##STR15##

N-Boc-4-Acetylpiperidine (4-2)

To a stirred suspension of 4-1 (5.2 g, 32 mmol), NEt₃ (5.3 mL, 38 mmol),and DMF (100 mL) at OOC was added Boc₂ O followed by removal of thecooling bath. After 20 hr, the reaction mixture was diluted with EtOAcand then washed with H₂ O, sat. NaHCO₃, 5% KHSO₄, and brine,dried(MgSO₄), and concentrated. Flash chromatography (silica, 30%EtOAc/hexanes) gave 4-2 as a colorless oil.

TLC Rf=0.31 (silica, 30% EtOAc/hexanes), ¹ H NMR (300 MHz, CDCl₃) δ4.08(bs, 2H), 2.80 (m, 2H), 2.45 (m, 1H), 2.17 (s, 3H), 1.83 (m, 2H), 1.48(m, 2H), 1.46 (s, 9H).

N-Boc-4-( 1.8!-Naphthyridin-2-yl)piperidine (4-3)

A mixture of 4-2 (4.0 g, 17,6 mmol), 1-6 (2.1 g, 17.6 mmol), L-proline(0.3 g, 2.6 mmol), and ethanol (88 mL) was refluxed for 40 hr. Thecooled reaction mixture was concentrated and then purified by flashchromatography (silica, 30% EtOAc/hexanes to 3% isopropanol/EtOAc) togive 4-3 as a yellow solid.

TLC Rf=0.29 (silica, EtOAc), ¹ H NMR (300 MHz, CDCl₃) δ9.10 (m, 1H),8.18 (m 1H), 8.11 (d, J=8 Hz, 1H), 7.46 (m, 1H), 7.41 (d, J=8 Hz, 1H),4.30 (m, 2H), 3.14 (m, 1H), 2.90 (m, 2H), 2.00 (m, 4H), 1.45 (s, 9H).

N-Boc-4-(5,6,7,8-Tetrahydro- 1,8!naphthyridin-2-yl)piperidine (4-4)

A mixture of 4-3 (2.6 g, 8.4 mmol), 10% Pd/C (0.52 g), and EtOAc (100mL) was stirred under a hydrogen atmosphere (1 atm) for 20 hr. Thereaction mixture was then filtered through a celite pad and the filtrateconcentrated. Flash chromatography (silica, 70% hexanes/EtOAc to EtOAc)gave 4-4 as a colorless oil.

TLC Rf=0.15 (silica, 10% isopropanol/EtOAc), ¹ H NMR (300 MHz, CDCl₃)δ7.13 (d, J=8 Hz, 1H), 6.33 (d, J=8 Hz, 1H), 4.20 (m, 2H), 3.42 (m, 2H),2.79 (m, 2H), 2.70 (m, 2H), 2.62 (m, 1H), 1.90 (m, 4H), 1.60 (m, 2H),1.43 (s, 9H).

4-(5,6,7,8-Tetrahydro- 1,8!naphthyridin-2-yl)piperidine.2 HCl (4-5)

HCl gas was bubbled through a solution of 4-4 (1.6 g, 4.8 mmol) andEtOAc (40 mL) at 0° C. for 3 min. The solution was stirred for anadditional 1 hr followed by concentration to give 4-5 as a yellow foam.

¹ H NMR (300 MHz, CD₃ OD) δ7.63 (d, J=8 Hz, 1H), 6.67 (d, J=8 Hz, 1H),3.52 (m, 4H), 3.16 (m, 2H), 3.07 (m, 1H), 2.83 (d, J=6 Hz, 2H), 2.20 (m,2), 1.96 (m, 4H).

Ethyl 4-(5,6,7,8-tetrahydro- 1,8!naphthyridin-2-yl)piperidinyl-malonate(4-6)

A mixture of 4-5 (150 mg, 0.52 mmol), ethyl hydrogen malonate (76 mg,0.57 mmol), HOBT (70 mg, 0.52 mmol), NMM (0.29 mL, 2.1 mmol), and DMF (4mL) was treated with EDC (97 mg, 0.52 mmol). After 20 hr, the reactionmixture was diluted with EtOAc and then washed with sat. NaHCO₃, H₂ O,and brine, dried (MgSO₄), and concentrated. Flash chromatography(silica, EtOAc) gave 4-6 as an oil.

¹ H NMR (300 MHz, CDCl₃) δ7.05 (m, 1H), 6.35 (d, J=8 Hz, 1H), 4.78 (bs,1H), 4.70 (m, 1H), 4.50 (m, 1H), 4.20 (m, 2H), 3.76 (m, 2H), 3.50 (m,3H), 3.40 (m, 2H), 3.30 (m, 1H), 2.85-2.50 (m, 4H), 2.10-1.50 (m, 6H),1.28 (m, 3H).

4-(5,6,7,8-Tetrahydro-1,8!naphthyridin-2-yl)piperidinyl-malonyl-3(S)-pyridin-3-yl-β-alanineethyl ester (4-7)

A solution of 4-6 (43 mg, 0.13 mmol), 1N NaOH (0.32 mL), THF (1 mL), andH₂ O (1 mL) was stirred at ambient temperature for 1 hr. The reactionmixture was then treated with 1N HCl (0.32 mL) and concentrated to givea semi-solid. The residue was dissolved in DMF (4 mL) and treatedsequentially with 1-11(37 mg, 0.14 mmol), HOBT (19 mg, 0.14 mmol), NMM(138 uL, 0.98 mmol), and EDC (27 mg, 0.14 mmol). After 20 hr, thereaction mixture was diluted with EtOAc and then washed with sat,NaHCO₃, H₂ O, and brine, dried (MgSO₄), and concentrated. Flashchromatography (silica, 70:15:5 CHCl₃ /methanol/EtOAc) gave 4-7 as ayellowish oil.

¹ H NMR (300 MHz, CDCl₃) δ9.02 (d, J=8 Hz, 0.5H), 8.90 (d, J=8 Hz,0.5H), 8.62 (m, 1H), 8.50 (m, 1H), 7.70 (m, 1H), 7.23 (m, 1H), 7.07 (d,J=8 Hz, 1H), 6.28 (m, 1H), 5.49 (m, 1H), 4.88 (m, 1H), 4.72 (m, 0.5 H),4.45(m, 0.5H), 4.10 (m, 2H), 3.90 (m, 1H), 3.40 (m, 4H), 3.00-2.50 (m,6H), 2.05-1.50 (m, 6H), 1.17 (t, J=7 Hz, 3H)

4-(5,6,7,8-Tetrahydro-1,8!naphthyridin-2-yl)piperidinyl-malonyl-3(S)-pyridin-3-yl-β-alanine(4-8)

A mixture of 4-7 (20 mg, 42 umol), 1N NaOH (104 uL), THF (0.2 mL), andH₂ O (0.2 mL) was stirred at ambient temperature for 3 hr. The reactionmixture was treated with 1N HCl (110 uL) and then concentrated. FlashChromatography (silica, 15:15:1:1 EtOAc/ethanol/H₂ O/NH₄ OH) gave 4-8 asa white solid.

TLC Rf=0.15 (silica, 15:10:1:1 EtOAc/ethanol/H₂ O/NH₄ OH), ¹ H NMR (300MHz, CD₃ OD) δ8.58 (m, 1H), 8.42 (m, 1H), 7.86 (m, 1H), 7.42 (m, 2H),6.50 (m, 1H), 5.40 (m, 1H), 4.48 (m, 1H), 3.88 (m, 1H), 3.70-3.00 (m,5H), 3.00-2.40 (m, 6H), 2.00-1.00 (m, 6H).

Utilizing the chemistry contained in Schemes 1-8, the followingcompounds 5, 6, 7 and 8 are prepared. ##STR16##

N-(4-Iodo-phenylsulfonylamino )-L-asparagine (9-2)

To a stirred solution of acid 9-1(4.39 g, 33.2 mmol), NaOH (1.49 g, 37.2mmol), dioxane (30 ml) and H₂ O (30 ml) at 0° C. was added pipsylchloride (10.34 g, 34.2 mmol). After ˜5 minutes, NaOH (1.49, 37.2 mmol)dissolved in 15 ml H₂ O, was added followed by the removal of thecooling bath. After 2.0 h, the reaction mixture was concentrated. Theresidue was dissolved in H₂ O (300 ml) and then washed with EtOAc. Theaqueous portion was cooled to 0° C. and then acidified with concentratedHCl. The solid was collected and then washed with Et₂ O to provide acid9-2 as a white solid.

¹ H NMR (300 MHz, D₂ O) δ7.86 (d, 2H, J=8 Hz), 7.48 (d, 2H, J=8 Hz) 3.70(m, 1H), 2.39 (m, 2H).

2(S)-(4-Iodo-phenylsulfonylamino)-β-alanine (9-3)

To a stirred solution of NaOH (7.14 g, 181.8 mmol) and H₂ O (40 ml) at0° C. was added Br₂ (1.30 ml, 24.9 mmol) dropwise over a ten minuteperiod. After ˜5 minutes, acid 9-2 (9.9 g, 24.9 mmol), NaOH (2.00 g,49.8 mmol) and H₂ O (35 ml) were combined, cooled to 0° C. and thenadded in a single portion to the reaction. After stirring for 20 minutesat 0° C., the reaction was heated to 90° C. for 30 minutes and thenrecooled to 0° C. The pH was adjusted to ˜7 by dropwise addition ofconcentrated HCl. The solid was collected, washed with EtOAc, and thendried in vacuo to provide acid 9-3 as a white solid.

¹ H NMR (300 MHz, D₂ O) δ8.02 (d, 2H, J=8 Hz), 7.63 (d, 2H, J=8 Hz),4.36 (m, 1H), 3.51 (dd, 1H, J=5 Hz, 13 Hz) 3.21 (m, 1H).

Ethyl 2(S)-(4-iodo-phenylsulfonylamino)-β-alanine-hydrochloride (9-4)

HCl gas was rapidly bubbled through a suspension of acid 9-3 (4.0 g,10.81 mmol) in EtOH (50 ml) at 0° C. for 10 minutes. The cooling bathwas removed and the reaction was heated to 60° C. After 18 h, thereaction was concentrated to provide ester 9-4 as a white solid.

¹ H NMR (300 MHz, CD₃ OD) δ7.98 (d, 2H, J=8 Hz), 7.63 (d, 2H, J=8 Hz),4.25 (q, 1H, J=5 Hz), 3.92 (m, 2H), 3.33 (m, 1H), 3.06 (m, 1H), 1.01 (t,3H, J=7 Hz).

Ethyl 4- 2-(2-Aminopyridin-6-yl)ethyl!benzoate (9-5)

A mixture of ester 9-5a (700 mg, 2.63 mmol), (for preparation, see:Scheme 29 of PCT International Application Publication No. WO 95/32710,published Dec. 7, 1995) 10% Pd/C (350 mg) and EtOH were stirred under 1atm H₂. After 20 h, the reaction was filtered through a celite pad andthen concentrated to provide ester 9-5 as a brown oil.

TLC R_(f) =0.23 (silica, 40% EtOAc/hexanes) ¹ H NMR (300 MHz, CDCl₃)δ7.95 (d, 2H, J=8 Hz), 7.26 (m, 3H), 6.43 (d, 1H, J=7 Hz), 6.35 (d, 1H,J=8 Hz), 4.37 (m, 4H), 3.05 (m, 2H), 2.91 (m, 2H), 1.39 (t, 3H, J=7 Hz).

4- 2-(2-Aminopyridin-6-yl)ethyl!benzoic acid hydrochloride (9-6)

A suspension of ester 9-5 (625 mg, 2.31 mmol) in 6N HCl (12 ml) washeated to 60° C. After ˜20 h, the reaction was concentrated to give acid9-6 as a tan solid.

¹ H NMR (300 MHz, CD₃ OD) δ7.96 (d, 2H, J=8 Hz), 7.80 (m, 1H), 7.33 (d,2H, J=8 Hz), 6.84 (d, 1H, J=9 Hz), 6.69 (d, 1H, J=7 Hz), 3.09 (m, 4H).

Ethyl 4-2-(2-Aminopyridin-6-yl)ethyl!benzoyl-2(S)-(4-iodophenylsulfonylamino)-.beta.-alanine(9-7)

A solution of acid 9-6 (400 mg, 1.43 mmol), amine 94 (686 mg, 1.57mmol), EDC (358 mg, 1.86 mmol), HOBT (252 mg, 1.86 mmol), NMM (632 μl,5.72 mmol) and DMF (10 ml) was stirred for ˜20 h. The reaction wasdiluted with EtOAc and then washed with sat NaHCO₃, brine, dried (MgSO₄)and concentrated. Flash chromatography (silica, EtOAC AE 5%isopropanol/EtOAc) provided amide 9-7 as a white solid.

TLC R_(f) =0.4 (silica, 10% isopropanol/EtOAc) ¹ H NMR (300 MHz, CD₃ OD)δ7.79 (d, 2H, J=9 Hz) 7.61 (d, 2H, J=8 Hz), 7.52 (d, 2H, J=9 Hz), 7.29(m, 1H), 7.27 (d, 2H, J=8 Hz), 4.20 (m, 1H), 3.95 (q, 2H, J=7 Hz), 3.66(dd, 1H, J=6 Hz, 14 Hz), 3.49 (dd, 1H, J=8Hz, 13 Hz), 3.01 (m, 2H), 2.86(m, 2H), 1.08 (t, 3H, J=7 Hz).

4-2-(2-Aminopyridin-6-yl)ethyl!benzoyl-2(S)-(4-iodophenylsulfonylamino)-.beta.-alanine(9-8)

A solution of ester 9-7 (200 mg, 0.3213 mmol) and 6N HCl (30 ml) washeated to 60° C. After ˜20 h, the reaction mixture was concentrated.Flash chromatography (silica, 20:20:1:1 EtOAc/EtOH/NH₄ OH/H₂ O) providedacid 9-8 as a white solid.

TLC R_(f) =0.45 (silica, 20:20:1:1 EtOAc/EtOH/NH₄ OH/H₂ O) ¹ H NMR (400MHz, DMSO) δ8.40 (m, 1H), 8.14 (Bs, 1H), 7.81 (d, 2H, J=8 Hz), 7.62 (d,2H, J=8 Hz), 7.48 (d, 2H, J=8 Hz), 7.27 (m, 3H), 6.34 (d, 1H, J=7 Hz),6.25 (d, 1H, J=8 Hz), 5.85 (bs, 2H), 3.89 (bs, 1H), 3.35 (m, 2H), 2.97(m, 2H), 2.79 (m, 2H).

4-2-(2-Aminopyridin-6-yl)ethyl)benzoyl-2(S)-(4-trimethylstannyl-phenylsulfonylamino-β-alanine(9-9)

A solution of iodide 9-8 (70 mg, 0.1178 mmol), (CH₃ Sn)₂ (49 μl, 0.2356mmol), Pd(PPh₃)₄ (5 mg) and dioxane (7 ml) was heated to 90° C. After 2h, the reaction was concentrated and then purified by prep HPLC(Delta-Pak C₁₈ 15 μM 100 A°, 40×100 mm; 95:5 AE 5:95 H₂ O/CH₃ CN)provided the trifluoroacetate salt. The salt was suspended in H₂ O (10ml), treated with NH₄ OH (5 drops) and then lyophilized to provide amide9-9 as a white solid.

¹ H NMR (400 MHz, DMSO) δ8.40 (m, 1H), 8.18 (d, 1H, J=8 Hz), 7.67 (m,5H), 7.56 (d, 2H, J=8 Hz), 7.29 (d, 2H, J=8 Hz), 6.95-7.52 (m, 2H), 6.45(bs, 2H), 4.00 (m, 1H), 3.50 (m, 1H), 3.33 (m, 1H), 2.97 (m, 2H), 2.86(m, 2H).

4- 2-(2-Aminopyridin-6-yl)ethyl!benzoyl-2(S)-4-¹²⁵iodophenylsulfonylamino-β-alanine (9-10)

An iodobead (Pierce) was added to a shipping vial of 5 mCi of Na¹²⁵ I(Amersham, IMS30) and stirred for five minutes at room temperature. Asolution of 0.1 mg of 9-9 in 0.05 mL of 10% H₂ SO₄ /MeOH was made andimmediately added to the Na¹²⁵ I/iodobead vial. After stirring for threeminutes at room temperature, approximately 0.04-0.05 mL of NH₄ OH wasadded so the reaction mixture was at pH 6-7. The entire reaction mixturewas injected onto the HPLC for purification Vydac peptide-protein C-18column, 4.6×250 mm, linear gradient of 10% acetonitrile (0.1% (TFA):H₂ O(0.1% TFA) to 90% acetonitrile (0.1% TFA):H₂ O (0.1% TFA) over 30minutes, 1 mL/min!. The retention time of 9-10 is 17 minutes under theseconditions. Fractions containing the majority of the radioactivity werepooled, lyophilized and diluted with ethanol to give approximately 1 mCiof 9-10, which coeluted on HPLC analysis with an authentic sample of9-8.

Instrumentation: Analytical and preparative HPLC was carried out using aWaters 600E Powerline Multi Solvent Delivery System with 0.1 mL headswith a Rheodyne 7125 injector and a Waters 990 Photodiode Array Detectorwith a Gilson FC203 Microfraction collector. For analytical andpreparative HPLC a Vydac peptide-protein C-18 column, 4.6×250 mm wasused with a C-18 Brownlee modular guard column. The acetonitrile usedfor the HPLC analyses was Fisher Optima grade. The HPLC radiodetectorused was a Beckman 170 Radioisotope detector. A Vydac C-18 protein andpeptide column, 3.9×250 mm was used for analytical and preparative HPLC.Solutions of radioactivity were concentrated using a Speedvac vacuumcentrifuge. Calibration curves and chemical concentrations weredetermined using a Hewlett Packard Model 8452A UV/Vis Diode ArraySpectrophotometer. Sample radioactivities were determined in a PackardA5530 gamma counter.

The test procedures employed to measure avb3 binding and the boneresorption inhibiting activity of the compounds of the present inventionare described below.

BONE RESORPTION-PIT ASSAY

When osteoclasts engage in bone resorption, they will literally causethe formation of pits in the surface of bone that they are acting upon.Therefore, when testing compounds for their ability to inhibitosteoclasts, it is useful to measure the ability of osteoclasts toexcavate these resorption pits when the inhibiting compound is present.

Consecutive 200 micron thick cross sections from a six mm cylinder ofbovine femur diaphysis were cut with a low speed diamond saw (Isomet,Beuler, Ltd., Lake Bluff, Ill.). Bone slices were pooled, placed in a10% ethanol solution and refrigerated until further use.

Prior to experimentation, bone slices were ultrasonicated twice, 20minutes each in H₂ O. Cleaned slices were placed in 96 well plates suchthat two control lanes and one lane for each drug dosage are available.Each lane represents either triplicate or quadruplicate cultures. Thebone slices in 96 well plates were sterilized by UV irradiation. Priorto incubation with osteoclasts, the bone slices were hydrated by theaddition of 0.1 ml Medium 199, pH 6.9 containing 15% fetal bovine serumand 1% penicillin/streptomycin.

Osteoclasts were isolated from the long bones of 1 to 3 day old rat pups(Sprague-Dawley) by modifications of Chambers et al., (J. Cell. Science,66:383-399). The resulting suspension (0.75 ml/bone) was gentlytriturated 90-120 times using a wide bore transfer pipet. The cellularpopulation was separated from bone fragments by a cell strainer with a100 micron nylon mesh. 100 μl of the cell suspension was placed ontoeach bone slice. Test compounds were then added at the desiredexperimental concentrations.

Bone slices exposed to osteoclasts for 20-24 hrs were processed forstaining. Tissue culture media was removed from each bone slice. Eachwell was washed with 200 μl of H₂ O, and the bone slices were then fixedfor 20 minutes in 2.5% glutaraldehyde, 0.1 M cacodylate, pH 7.4. Afterfixation, any remaining cellular debris was removed by 2 min.ultrasonication in the presence of 0.25 M NH₄ OH followed by 2×15 minultrasonication in H₂ O. The bone slices were immediately stained for6-8 min with filtered 1% toluidine blue and 1% borax.

After the bone slices have dried, resorption pits were counted in testand control slices. Resorption pits were viewed in a Microphot Fx(Nikon) fluorescence microscope using a polarizing Nikon IGS filtercube. Test dosage results were compared with controls and resulting IC₅₀values were determined for each compound tested.

The appropriateness of extrapolating data from this assay to utility anduse in mammalian (including human) disease states is supported by theteaching found in Sato, M., et al., Journal of Bone and MineralResearch, Vol. 5, No. 1, 1990. That article teaches that certainbisphosphonates have been used clinically and appear to be effective inthe treatment of Paget's disease, hypercalcemia of malignancy,osteolytic lesions produced by bone metastases, and bone loss due toimmobilization or sex hormone deficiency. These same bisphosphonates arethen tested in the resorption pit assay described above to confirm acorrelation between their known utility and positive performance in theassay.

EIB ASSAY

Duong et al., J. Bone Miner. Res., 8:S 378, describe a system forexpressing the human integrin αvβ3. It has been suggested that theintegrin stimulates attachment of osteoclasts to bone matrix, sinceantibodies against the integrin, or RGD-containing molecules, such asechistatin (European Publication 382 451), can effectively block boneresorption.

Reaction Mixture:

1. 175 μl TBS buffer (50 mM Tris.HCl pH 7.2, 150 mM NaCl, 1% BSA, 1 mMCaC1₂, 1 mM MgCl₂).

2. 25 μl cell extract (dilute with 100 mM octylglucoside buffer to give2000 cpm/25 μl).

3. ¹²⁵ I-echistatin (25 μl/50,000 cpm) (see EP 382 451).

4. 25 μl buffer (total binding) or unlabeled echistatin (non-specificbinding).

The reaction mixture was then incubated for 1 h at room temp. Theunbound and the bound αvβ3 were separated by filtration using a SkatronCell Harvester. The filters (prewet in 1.5% poly-ethyleneimine for 10mins) were then washed with the wash buffer (50 mM Tris HCl, 1 mM CaCl₂/MgCl₂, pH 7.2). The filter was then counted in a gamma counter.

SPA ASSAY

Materials:

1. Wheatgerm agglutinin Scintillation Proximity Beads (SPA): Amersham

2. Octylglucopyranoside: Calbiochem

3. HEPES: Calbiochein

4. NaCl: Fisher

5. CaCl₂ : Fisher

6. MgCl₂ : SIGMA

7. Phenylmethylsulfonylfluoride (PMSF): SIGMA

8. Optiplate: PACKARD

9. 9-10 (specific activity 500-1000 Ci/mmole)

10. test compound

11. Purified integrin receptor: αvβ3 was purified from 293 cellsoverexpressing αvβ3 (Duong et al., J. Bone Min. Res., 8:S378, 1993)according to Pytela (Methods in Enzymology, 144:475, 1987)

12. Binding buffer: 50 mM HEPES, pH 7.8, 100 mM NaCl, 1 mM Ca²⁺ /Mg²⁺,0.5 mM PMSF

13. 50 mM octylglucoside in binding buffer: 50-OG buffer

Procedure:

1. Pretreatment of SPA beads:

500 mg of lyophilized SPA beads were first washed four times with 200 mlof 50-OG buffer and once with 100 ml of binding buffer, and thenresuspended in 12.5 ml of binding buffer.

2. Preparation of SPA beads and receptor mixture

In each assay tube, 2.5 μl (40 mg/ml) of pretreated beads were suspendedin 97.5 μl of binding buffer and 20 ml of 50-OG buffer. 5 μl (˜30 ng/ml)of purified receptor was added to the beads in suspension with stirringat room temperature for 30 minutes. The mixture was then centrifuged at2,500 rpm in a Beckman GPR Benchtop centrifuge for 10 minutes at 4° C.The pellets were then resuspended in 50 μl of binding buffer and 25 μlof 50-OG buffer.

3. Reaction

The following were sequentially added into Optiplate in correspondingwells:

(i) Receptor/beads mixture (75 μl)

(ii) 25 μl of each of the following: compound to be tested, bindingbuffer for total binding or 9-8 for non-specific binding (finalconcentration 1 μM)

(iii) 9-10 in binding buffer (25 μl, final concentration 40 pM)

(iv) Binding buffer (125 μl)

(v) Each plate was sealed with plate sealer from PACKARD and incubatedovernight with rocking at 4° C.

4. Plates were counted using PACKARD TOPCOUNT

5. 1% inhibition was calculated as follows:

A=total counts

B=nonspecific counts

C=sample counts

% inhibition= {(A-B)-(C-B)}/(A-B)!/(A-B)×100

OCFORM ASSAY

Osteoblast-like cells (1.8 cells), originally derived from mousecalvaria, were plated in CORNING 24 well tissue culture plates in a MEMmedium containing ribo- and deoxyribonucleosides, 10% fetal bovine serumand penicillin-streptomycin. Cells were seeded at 40,000/well in themorning. In the afternoon, bone marrow cells were prepared from six weekold male Balb/C mice as follows:

Mice were sacrificed, tibiae removed and placed in the above medium. Theends were cut off and the marrow was flushed out of the cavity into atube with a 1 mL syringe with a 27.5 gauge needle. The marrow wassuspended by pipetting up and down. The suspension was passedthrough >100 μm nylon cell strainer. The resulting suspension wascentrifuged at 350×g for seven minutes. The pellet was resuspended, anda sample was diluted in 2% acetic acid to lyse the red cells. Theremaining cells were counted in a hemacytometer. The cells were pelletedand resuspended at 1×10⁶ cells/mL. 50 μL was added to each well of 1.8cells to yield 50,000 cells/well and 1,25-dihydroxy-vitamin D₃ (D₃) wasadded to each well to a final concentration of 10 nM. The cultures wereincubated at 37° C. in a humidified, 5% CO₂ atmosphere. After 48 h, themedium was changed. 72 h after the addition of bone marrow, testcompounds were added with fresh medium containing D₃ to quadruplicatewells. Compounds were added again after 48 h with fresh mediumcontaining D₃. After an additional 48 h the medium was removed, cellswere fixed with 10% formaldehyde in phosphate buffered saline for 10minutes at room temperature, followed by a 1-2 minute treatment withethanol:acetone (1:1) and air dried. The cells were then stained fortartrate resistant acid phosphatase as follows:

The cells were stained for 10-15 minutes at room temperature with 50 mMacetate buffer, pH 5.0 containing 30 mM sodium tartrate, 0.3 mg/mL FastRed Violet LB Salt and 0.1 mg/mL Naphthol AS -MX phosphate. Afterstaining, the plates were washed extensively with deionized water andair dried. The number of multinucleated, positive staining cells werecounted in each well.

Representative compounds of the present invention were tested and foundto bind to human αvβ3 integrin. These compounds were found to have IC₅₀values in the range of 1.0 to 10,000 nM in the SPA assay.

EXAMPLE OF A PHARMACEUTICAL FORMULATION

As a specific embodiment of an oral composition, 100 mg of compound 1-13is formulated with sufficient finely divided lactose to provide a totalamount of 580 to 590 mg to fill a size O hard gel capsule.

While the invention has been described and illustrated in reference tocertain preferred embodiments thereof, those skilled in the art willappreciate that various changes, modifications and substitutions can bemade therein without departing from the spirit and scope of theinvention. For example, effective dosages other than the preferred dosesas set forth hereinabove may be applicable as a consequence ofvariations in the responsiveness of the mammal being treated forseverity of bone disorders caused by resorption, or for otherindications for the compounds of the invention indicated above.Likewise, the specific pharmacological responses observed may varyaccording to and depending upon the particular active compound selectedor whether there are present pharmaceutical carriers, as well as thetype of formulation and mode of administration employed, and suchexpected variations or differences in the results are contemplated inaccordance with the objects and practices of the present invention. Itis intended, therefore, that the invention be limited only by the scopeof the claims which follow and that such claims be interpreted asbroadly as is reasonable.

What is claimed is:
 1. A compound of the formula ##STR17## wherein X isselected from a 5- or 6-membered monocyclic aromatic or nonaromatic ringsystem containing 1, 2, 3 or 4 heteroatoms selected from N, O or Swherein the 5- or 6-membered ring system is either unsubstituted orsubstituted with R¹ and R², ora 9- to 10-membered polycyclic ringsystem, wherein one or more of the rings is aromatic, and wherein thepolycyclic ring system contains 1, 2, 3 or 4 heteroatoms selected fromN, O or S, and wherein the polycyclic ring system is eitherunsubstituted or substituted with R¹ and R² ; Y is selected from##STR18## Z is absent or is a 4-11 membered aromatic or nonaromaticmono- or polycyclic ring system containing 0 to 6 double bonds, andcontaining 0 to 6 heteroatoms chosen from N, O and S, and wherein thering system is either unsubstituted or substituted on a carbon ornitrogen atom with one or more groups independently selected from R¹⁴,R¹⁵, R¹⁶ and R¹⁷ ; R¹, R², R³, R⁴, R⁵, R¹¹, R¹², R¹³, R¹⁶ and R¹⁷ areeach independently selected fromhydrogen, halogen, C₁₋₁₀ alkyl, C₃₋₈cycloalkyl, aryl, aryl C₁₋₈ alkyl, amino, amino C₁₋₈ alkyl, C₁₋₃acylamino, C₁₋₃ acylamino C₁₋₈ alkyl, C₁₋₆ alkylamino, C₁₋₆alkylamino-C₁₋₈ alkyl, C₁₋₆ dialkylamino, C₁₋₆ dialkylamino C₁₋₈ alkyl,C₁₋₄ alkoxy, C₁₋₄ alkoxy C₁₋₆ alkyl, hydroxycarbonyl, hydroxycarbonylC₁₋₆ alkyl, C₁₋₃ alkoxycarbonyl, C₁₋₃ alkoxycarbonyl C₁₋₆ alkyl,hydroxycarbonyl-C₁₋₆ alkyloxy, hydroxy or hydroxy C₁₋₆ alkyl; R₆, R⁷,R¹⁴ and R¹⁵ are each independently selected fromhydrogen, aryl, halogen,aryl-(CH₂)_(p) --, hydroxyl, C₁₋₈ alkylcarbonylamino, aryl C₁₋₅ alkoxy,C₁₋₅ alkoxycarbonyl, aminocarbonyl, C₁₋₈ alkylaminocarbonyl, C₁₋₆alkylcarbonyloxy, C₃₋₈ cycloalkyl, amino, C₁₋₆ alkylamino, amino C₁₋₆alkyl, arylaminocarbonyl, aryl C₁₋₅ alkylaminocarbonyl, aminocarbonyl,aminocarbonyl C₁₋₆ alkyl, hydroxycarbonyl, hydroxycarbonyl C₁₋₆ alkyl,C₁₋₈ alkyl, either unsubstituted or substituted, with one or more groupsselected from: halogen, hydroxyl, C₁₋₅ alkylcarbonylamino, aryl C₁₋₅alkoxy, C₁₋₅ alkoxycarbonyl, aminocarbonyl, C₁₋₅ alkylaminocarbonyl,C₁₋₅ alkylcarbonyloxy, C₃₋₈ cycloalkyl, oxo, amino, C₁₋₃ alkylamino,amino C₁₋₃ alkyl, arylaminocarbonyl, aryl C₁₋₅ alkylaminocarbonyl,aminocarbonyl, aminocarbonyl C₁₋₄ alkyl, hydroxycarbonyl, orhydroxycarbonyl C₁₋₅ alkyl, HC.tbd.C(CH₂)_(r) -- C₁₋₆alkyl-C.tbd.C(CH₂)_(r) --, C₃₋₇ cycloalkyl-C.tbd.C(CH₂)_(r) --,aryl-C.tbd.C(CH₂)_(r) --, C₁₋₆ alkylaryl-C.tbd.C(CH₂)_(r) --, H₂C═CH(CH₂)_(r) --, C₁₋₆ alkyl-CH═CH(CH₂)_(r) --, C₃₋₇cycloalkyl-CH═CH(CH₂)_(r) --, aryl-CH═CH(CH₂)_(r) --, C₁₋₆alkylaryl-CH═CH(CH₂)_(r) --, C₁₋₆ alkyl-SO₂ (CH₂)_(r) --, C₁₋₆alkylaryl-SO₂ (CH₂)_(r) --, C₁₋₆ alkoxy, aryl C₁₋₆ alkoxy, aryl C₁₋₆alkyl, C₁₋₆ alkylamino C₁₋₆ alkyl, arylamino, arylamino C₁₋₆ alkyl, arylC₁₋₆ alkylamino, aryl C₁₋₆ alkylamino C₁₋₆ alkyl, arylcarbonyloxy, arylC₁₋₆ alkylcarbonyloxy, C₁₋₆ dialkylamino, C₁₋₆ dialkylamino C₁₋₆ alkyl,C₁₋₆ alkylaminocarbonyloxy, C₁₋₈ alkylsulfonylamino, C₁₋₈alkylsulfonylamino C₁₋₆ alkyl, arylsulfonylamino C₁₋₆ alkyl, aryl C₁₋₆alkylsulfonylamino, aryl C₁₋₆ alkylsulfonylamino C₁₋₆ alkyl, C₁₋₈alkoxycarbonylamino, C₁₋₈ alkoxycarbonylamino C₁₋₈ alkyl,aryloxycarbonylamino C₁₋₈ alkyl, aryl C₁₋₈ alkoxycarbonylamino, arylC₁₋₈ alkoxycarbonylamino C₁₋₈ alkyl, C₁₋₈ alkylcarbonylamino, C₁₋₈alkylcarbonylamino C₁₋₆ alkyl, arylcarbonylamino C₁₋₆ alkyl, aryl C₁₋₆alkylcarbonylamino, aryl C₁₋₆ alkylcarbonylamino C₁₋₆ alkyl,aminocarbonylamino C₁₋₆ alkyl, C₁₋₈ alkylaminocarbonylamino, C₁₋₈alkylaminocarbonylamino C₁₋₆ alkyl, arylaminocarbonylamino C₁₋₆ alkyl,aryl C₁₋₈ alkylaminocarbonylamino, aryl C₁₋₈ alkylaminocarbonylaminoC₁₋₆ alkyl, aminosulfonylamino C₁₋₆ alkyl, C₁₋₈ alkylaminosulfonylamino,C₁₋₈ alkylaminosulfonylamino C₁₋₆ alkyl, arylaminosulfonylamino C₁₋₆alkyl, aryl C₁₋₈ alkylaminosulfonylamino, aryl C₁₋₈alkylaminosulfonylamino C₁₋₆ alkyl, C₁₋₆ alkylsulfonyl, C₁₋₆alkylsulfonyl C₁₋₆ alkyl, arylsulfonyl C₁₋₆ alkyl, aryl C₁₋₆alkylsulfonyl, aryl C₁₋₆ alkylsulfonyl C₁₋₆ alkyl, C₁₋₆ alkylcarbonyl,C₁₋₆ alkylcarbonyl C₁₋₆ alkyl, arylcarbonyl C₁₋₆ alkyl, aryl C₁₋₆alkylcarbonyl, aryl C₁₋₆ alkylcarbonyl C₁₋₆ alkyl, C₁₋₆alkylthiocarbonylamino, C₁₋₆ alkylthiocarbonylamino C₁₋₆ alkyl,arylthiocarbonylamino C₁₋₆ alkyl, aryl C₁₋₆ alkylthiocarbonylamino, arylC₁₋₆ alkylthiocarbonylamino C₁₋₆ alkyl, C₁₋₈ alkylaminocarbonyl C₁₋₆alkyl, arylaminocarbonyl C₁₋₆ alkyl, aryl C₁₋₈ alkylaminocarbonyl, oraryl C₁₋₈ alkylaminocarbonyl C₁₋₆ alkyl,wherein any of the alkyl groupsmay be unsubstituted or substituted with R¹¹ and R¹² ; and provided thatthe carbon atom to which R⁶ and R⁷ are attached is itself attached to nomore than one heteroatom; or R⁶ and R⁷, or R¹⁴ and R¹⁵ are combined toform oxo; R⁸ and R9 are each independently selected fromhydrogen, aryl,halogen, aryl-(CH₂)_(p) --, hydroxyl, C₁₋₈ alkylcarbonylamino, aryl C₁₋₅alkoxy, C₁₋₅ alkoxycarbonyl, aminocarbonyl, C₁₋₈ alkylaminocarbonyl,C₁₋₆ alkylcarbonyloxy, C₃₋₈ cycloalkyl, amino, C₁₋₆ alkylamino, aminoC₁₋₆ alkyl, arylaminocarbonyl, aryl C₁₋₅ alkylaminocarbonyl,aminocarbonyl, aminocarbonyl C₁₋₆ alkyl, hydroxycarbonyl,hydroxycarbonyl C₁₋₆ alkyl, C₁₋₈ alkyl, either unsubstituted orsubstituted, with one or more groups selected from: halogen, hydroxyl,C₁₋₅ alkylcarbonylamino, aryl C₁₋₅ alkoxy, C₁₋₅ alkoxycarbonyl,aminocarbonyl, C₁₋₅ alkylaminocarbonyl, C₁₋₅ alkylcarbonyloxy, C₃₋₈cycloalkyl, oxo, amino, C₁₋₃ alkylamino, amino C₁₋₃ alkyl,arylaminocarbonyl, aryl C₁₋₅ alkylaminocarbonyl, aminocarbonyl,aminocarbonyl C₁₋₄ alkyl, hydroxycarbonyl, or hydroxycarbonyl C₁₋₅alkyl, HC.tbd.C(CH₂)_(r) -- C₁₋₆ alkyl-C.tbd.C(CH₂)_(r) --, C₃₋₇cycloalkyl-C.tbd.C(CH₂)_(r) --, aryl-C.tbd.C(CH₂)_(r) --, C₁₋₆alkylaryl-C.tbd.C(CH₂)_(r) --, H₂ C═CH(CH₂)_(r) --, C₁₋₆alkyl-CH═CH(CH₂)_(r) --, C₃₋₇ cycloalkyl-CH═CH(CH₂)_(r) --,aryl-CH═CH(CH₂)_(r) --, C₁₋₆ alkylaryl-CH═CH(CH₂)_(r) --, C₁₋₆ alkyl-SO₂(CH₂)_(r) --, C₁₋₆ alkylaryl-SO₂ (CH₂)_(r) --, C₁₋₆ alkoxy, aryl C₁₋₆alkoxy, aryl C₁₋₆ alkyl, C₁₋₆ alkylamino C₁₋₆ alkyl, arylamino,arylamino C₁₋₆ alkyl, aryl C₁₋₆ alkylamino, aryl C₁₋₆ alkylamino C₁₋₆alkyl, arylcarbonyloxy, aryl C₁₋₆ alkylcarbonyloxy, C₁₋₆ dialkylamino,C₁₋₆ dialkylamino C₁₋₆ alkyl, C₁₋₆ alkylaminocarbonyloxy, C₁₋₈alkylsulfonylamino, C₁₋₈ alkylsulfonylamino C₁₋₆ alkyl,arylsulfonylamino C₁₋₆ alkyl, aryl C₁₋₆ alkylsulfonylamino, aryl C₁₋₆alkylsulfonylamino C₁₋₆ alkyl, C₁₋₈ alkoxycarbonylamino, C₁₋₈alkoxycarbonylamino C₁₋₈ alkyl, aryloxycarbonylamino C₁₋₈ alkyl, arylC₁₋₈ alkoxycarbonylamino, aryl C₁₋₈ alkoxycarbonylamino C₁₋₈ alkyl, C₁₋₈alkylcarbonylamino, C₁₋₈ alkylcarbonylamino C₁₋₆ alkyl,arylcarbonylamino C₁₋₆ alkyl, aryl C₁₋₆ alkylcarbonylamino, aryl C₁₋₆alkylcarbonylamino C₁₋₆ alkyl, aminocarbonylamino C₁₋₆ alkyl, C₁₋₈alkylaminocarbonylamino, C₁₋₈ alkylaminocarbonylamino C₁₋₆ alkyl,arylaminocarbonylamino C₁₋₆ alkyl, aryl C₁₋₈ alkylaminocarbonylamino,aryl C₁₋₈ alkylaminocarbonylamino C₁₋₆ alkyl, aminosulfonylamino C₁₋₆alkyl, C₁₋₈ alkylaminosulfonylamino, C₁₋₈ alkylaminosulfonylamino C₁₋₆alkyl, arylaminosulfonylamino C₁₋₆ alkyl, aryl C₁₋₈alkylaminosulfonylamino, aryl C₁₋₈ alkylaminosulfonylamino C₁₋₆ alkyl,C₁₋₆ alkylsulfonyl, C₁₋₆ alkylsulfonyl C₁₋₆ alkyl, arylsulfonyl C₁₋₆alkyl, aryl C₁₋₆ alkylsulfonyl, aryl C₁₋₆ alkylsulfonyl C₁₋₆ alkyl, C₁₋₆alkylcarbonyl, C₁₋₆ alkylcarbonyl C₁₋₆ alkyl, arylcarbonyl C₁₋₆ alkyl,aryl C₁₋₆ alkylcarbonyl, aryl C₁₋₆ alkylcarbonyl C₁₋₆ alkyl, C₁₋₆alkylthiocarbonylamino, C₁₋₆ alkylthiocarbonylamino C₁₋₆ alkyl,arylthiocarbonylamino C₁₋₆ alkyl, aryl C₁₋₆ alkylthiocarbonylamino, arylC₁₋₆ alkylthiocarbonylamino C₁₋₆ alkyl, C₁₋₈ alkylaminocarbonyl C₁₋₆alkyl, arylaminocarbonyl C₁₋₆ alkyl, aryl C₁₋₈ alkylaminocarbonyl, arylC₁₋₈ alkylaminocarbonyl C₁₋₆ alkyl, C₇₋₂₀ polycyclyl C₀₋₈alkylsulfonylamino C₀₋₆ alkyl, C₇₋₂₀ polycyclyl C₀₋₈ alkylcarbonylaminoC₀₋₆ alkyl, C₇₋₂₀ polycyclyl C₀₋₈ alkylaminosulfonyolamino C₀₋₆ alkyl,C₇₋₂₀ polycyclyl C₀₋₈ alkylaminocarbonylamino C₀₋₆ alkyl, or C₇₋₂₀polycyclyl C₀₋₈ alkyloxycarbonylamino C₀₋₆ alkylwherein any of the alkylgroups may be unsubstituted or substituted with R¹¹ and R¹², wherein anyof the polycyclyl may be unsubstituted or substituted with R¹⁸, R¹⁹, R²⁰and R²¹, and provided that the carbon atom to which R⁸ and R⁹ areattached is itself attached to no more than one heteroatom; or R⁸ and R⁹are combined to form oxo; R¹⁰ is selected fromhydrogen, C₁₋₈ alkyl,aryl, aryl C₁₋₈ alkyl, aryl C₁₋₆ alkoxy, C₁₋₈ alkylcarbonyloxy C₁₋₄alkyl, aryl C₁₋₈ alkylcarbonyloxy C₁₋₄ alkyl, C₁₋₈alkylaminocarbonylmethylene, or C₁₋₈ dialkylaminocarbonylmethylene; R¹⁸,R¹⁹, R²⁰ and R²¹ are each independently selected fromhydrogen, halogen,C₁₋₁₀ alkyl, C₃₋₈ cycloalkyl, oxo, aryl, aryl C₁₋₈ alkyl, amino, aminoC₁₋₈ alkyl, C₁₋₃ acylamino, C₁₋₃ acylamino C₁₋₈ alkyl, C₁₋₆ alkylamino,C₁₋₆ alkylamino-C₁₋₈ alkyl, C₁₋₆ dialkylamino, C₁₋₆ dialkylamino C₁₋₈alkyl, C₁₋₄ alkoxy, C₁₋₄ alkoxy C₁₋₆ alkyl, hydroxycarbonyl,hydroxycarbonyl C₁₋₆ alkyl, C₁₋₃ alkoxycarbonyl, C₁₋₃ alkoxycarbonylC₁₋₆ alkyl, hydroxycarbonyl-C₁₋₆ alkyloxy, hydroxy, hydroxy C₁₋₆ alkyl,C₁₋₆ alkyloxy-C₁₋₆ alkyl, nitro, cyano, trifluoromethyl,trifluoromethoxy, trifluoroethoxy, C₁₋₈ alkyl-S(O)_(q), C₁₋₈alkylaminocarbonyl, C₁₋₈ dialkylaminocarbonyl, C₁₋₈alkyloxycarbonylamino, C₁₋₈ alkylaminocarbonyloxy or C₁₋₈alkylsulfonylamino; m, n and r are each independently an integer from 0to 3; p is an integer from 1 to 4; and q is an integer from 0 to 2;andthe pharmaceutically acceptable salts thereof.
 2. The compound of claim1, whereinX isa 9- to 10-membered polycyclic ring system, wherein one ormore of the rings is aromatic, and wherein the polycyclic ring systemcontains 1, 2, 3 or 4 heteroatoms selected from N, O or S, and whereinthe polycyclic ring system is either unsubstituted or substituted withR¹ and R² ; andand the pharmaceutically acceptable salts thereof.
 3. Thecompound of claim 2 wherein the compound has the formula ##STR19##wherein X is selected from ##STR20## Z is absent or is selected from##STR21## R⁶ is selected from hydrogen,aryl, --(CH₂)_(p) -aryl, C₁₋₈alkyl, either unsubstituted or substituted, with one or more groupsselected from: halogen, hydroxyl, C₁₋₅ alkylcarbonylamino, aryl C₁₋₅alkoxy, C₁₋₅ alkoxycarbonyl, aminocarbonyl, C₁₋₅ alkylaminocarbonyl,C₁₋₅ alkylcarbonyloxy, C₃₋₈ cycloalkyl, oxo, amino, C₁₋₃ alkylamino,amino C₁₋₃ alkyl, arylaminocarbonyl, aryl C₁₋₅ alkylaminocarbonyl,aminocarbonyl, aminocarbonyl C₁₋₄ alkyl, hydroxycarbonyl, orhydroxycarbonyl C₁₋₅ alkyl, HC.tbd.C(CH₂)_(r) -- C₁₋₆alkyl-C.tbd.C(CH₂)_(r) --, C₃₋₇ cycloalkyl-C.tbd.C(CH₂)_(r) --,aryl-C.tbd.C(CH₂)_(r) --, C₁₋₆ alkylaryl-C.tbd.C(CH₂)_(r) --, H₂C═CH(CH₂)_(r) --, C₁₋₆ alkyl-CH═CH(CH₂)_(r) --, C₃₋₇cycloalkyl-CH═CH(CH₂)_(r) --, aryl-CH═CH(CH₂)_(r) --, C₁₋₆alkylaryl-CH═CH(CH₂)_(r) --, C₁₋₆ alkyl-SO₂ (CH₂)_(r) --, or C₁₋₆alkylaryl-SO₂ (CH₂)_(r) --,and the pharmaceutically acceptable saltsthereof.
 4. The compound of claim 3 of the formula ##STR22## wherein Yis selected from ##STR23## Z is absent or is selected from ##STR24## R⁶is selected from hydrogen, ##STR25## --(CH₂)_(p) indolyl,HC.tbd.C(CH₂)_(r) --C₁₋₆ alkyl-C.tbd.C(CH₂)_(r) --, C₃₋₇cycloalkyl-C.tbd.C(CH₂)_(r) --, aryl-C.tbd.C(CH₂)_(r) --, C₁₋₆alkylaryl-C═C(CH₂)_(r) --, H₂ C═CH(CH₂)_(r) --, C₁₋₆alkyl-CH═CH(CH₂)_(r) --, C₃₋₇ cycloalkyl-CH═CH(CH₂)_(r) --,aryl-CH═CH(CH₂)_(r) --, C₁₋₆ alkylaryl-CH=CH(CH₂)_(r) --, C₁₋₆ alkyl-SO₂(CH₂)_(r) --, or C₁₋₆ alkylaryl-SO₂ (CH₂)_(r) --; and R¹⁰ is selectedfrom hydrogen or C₁₋₈ alkyl;and the pharmaceutically acceptable saltsthereof.
 5. The compound of claim 4 selected fromEthyl3(S)-pyridin-3-yl-3-{2- 3-(5,6,7,8-tetrahydro-1,8!naphthyridin-2-yl)propylcarbamoyl!acelamino}propionate;3(S)-pyridin-3-yl-3-{2- 3-(5,6,7,8-tetrahydro- 1,8!naphthyridin-2-5yl)propylcarbamoyl!acetylamino}propionic acid; 3-(5,6,7,8-Tetrahydro-1,8!naphthyridin-2-ylmethyl)piperidinyl-malonyl-3(S)-pyridin-3-yl-β-alanineethyl ester; 3-(5,6,7,8-Tetrahydro-1,8!naphthyridin-2-ylmethyl)piperidinyl-malonyl-3(S)-pyridin-3-yl-β-alanine;4-(5,6,7,8-Tetrahydro-1,8!naphthyridin-2-ylmethyl)piperidinyl-malonyl-3(S)-pyridin-3-yl-β-alanineethyl ester; 4-(5,6,7,8-Tetrahydro-1,8!naphthyridin-2-ylmethyl)piperidinyl-malonyl-3(S)-pyridin-3-yl-β-alanine;4-(5,6,7,8-Tetrahydro-1,8!naphthyridin-2-yl)piperidinyl-malonyl-3(S)-pyridin-3-yl-β-alanineethyl ester; or 4-(5,6,7,8-Tetrahydro-1,8!naphthyridin-2-yl)piperidinyl-malonyl-3(S)-pyridin-3-yl-β-alanine;andthe pharmaceutically acceptable salts thereof.
 6. The compound of claim5 selected from3(S)-Pyridin-3-yl-3-{2- 3-(5,6,7,8-tetrahydro-1,8!naphthyridin-2-yl)propylcarbamoyl!acetylamino}propionic acid;3-(5,6,7,8-Tetrahydro-1,8!naphthyridin-2-ylmethyl)piperidinyl-malonyl-3(S)-pyridin-3-yl-β-alanine;4-(5,6,7,8-Tetrahydro-1,8!naphthyridin-2-ylmethyl)piperidinyl-malonyl-3(S)-pyridin-3-yl-β-alanine;or 4-(5,6,7,8-Tetrahydro-1,8!naphthyridin-2-yl)piperidinyl-malonyl-3(S)-pyridin-3-yl-β-alanine;andthe pharmaceutically acceptable salts thereof.
 7. A pharmaceuticalcomposition comprising the compound of claim 1 and a pharmaceuticallyacceptable carrier.
 8. A process for making a pharmaceutical compositioncomprising combining a compound of claim 1 and a pharmaceuticallyacceptable carrier.
 9. A method of eliciting a vitronectin antagonizingeffect in a mammal in need thereof, comprising administering to themammal a therapeutically effective amount of the compound of claim 1.10. The method of claim 9, wherein the vitronectin antagonizing effectis selected from inhibition of bone resorption, inhibition ofrestenosis, inhibition of angiogenesis, inhibition of diabeticretinopathy, inhibition of macular degeneration or inhibition of tumorgrowth.
 11. The method of claim 10, wherein the vitronectin antagonizingeffect is the inhibition of bone resorption.
 12. A method of treating orpreventing a condition mediated by antagonism of a vitronectin receptorin a mammal in need thereof, comprising administering to the mammal atherapeutically effective amount of the compound of claim
 1. 13. Themethod of claim 12, wherein the condition is elected from the groupconsisting of osteoporosis and cancer.
 14. The method of claim 13,wherein the condition is steoporosis.
 15. A method of inhibiting boneresorption in a mammal in need thereof, comprising administering to themammal a therapeutically effective amount of the compound of claim 1.16. A method of treating osteoporosis in a mammal in need thereof,comprising administering to the mammal a therapeutically effectiveamount of the compound of claim
 1. 17. A method of preventingosteoporosis in a mammal in need thereof, comprising administering tothe mammal a therapeutically effective amount of the compound ofclaim
 1. 18. A method of eliciting a vitronectin antagonizing effect ina mammal in need thereof, comprising administering to the mammal atherapeutically effective amount of the composition of claim
 7. 19. Amethod of treating or preventing a condition mediated by antagonism of avitronectin receptor in a mammal in need thereof, comprisingadministering to the mammal a therapeutically effective amount of thecomposition of claim
 7. 20. A method of inhibiting bone resorption in amammal in need thereof, comprising administering to the mammal atherapeutically effective amount of the composition of claim
 7. 21. Amethod of treating osteoporosis in a mammal in need thereof, comprisingadministering to the mammal a therapeutically effective amount of thecomposition of claim
 7. 22. A method of preventing osteoporosis in amammal in need thereof, comprising administering to the mammal atherapeutically effective amount of the composition of claim
 7. 23. Amethod of treating tumor growth in a mammal in need thereof, comprisingadministering to the mammal a therapeutically effective amount of acompound of claim 1 and one or more agents known to be cytotoxic orantiproliferative.